ES2299888T3 - PIRID (2,3-D) PYRIMIDIN-2,4-DIAMINS AS PDE INHIBITORS 2. - Google Patents

Abstract

A compound of formula (I) (See formula) and the salts of said pharmaceutically acceptable compounds, wherein: R 1 and R 2 are hydrogen or methoxy, with the proviso that R 1 and R 2 are not both hydrogen or both methoxy ; n is 1, 2, 3, or 4; X is a link; OR; S; C = O; -N (R) -; where R is hydrogen or (C1-C3) alkyl; -C (OH) -; or -SO2; and Y is benzoxazolyl; benzothiazolyl; benzofurazanyl; benzofuranyl; benzothiadiazolyl; bencisoxazolyl; bencisothiazolyl; benzimidazolyl; pyridyl; isatinyl; oxindolyl; indazolyl; indolyl; phenyl; thienyl; or furanyl; where Y is optionally substituted independently with between one and three halogen; trifluoromethyl; methoxy; -C (= O) CH3; cyano; -C (CH3) 2OH; -CH (CH3) OH; -CH (CF3) OH; -C (C = O) CF3; -SO2NH2; -C (= O) OCH3; -CH2COOH; (See formula) thiazolyl; or oxadiazolyl.

Description

Pyrido [2,3-d] pyrimidin-2,4-diamines as PDE 2 inhibitors.

Field of the Invention

The invention relates to certain pyrido [2,3-d] pyrimidin-2,4-diamines useful as PDE 2 inhibitors; the formulations of themselves; the combinations thereof; and the uses of same.

Background of the invention

The phosphodiesterase (PDE) enzyme family regulates the intracellular levels of cAMP or cGMP secondary messengers by hydrolytic control. Type II phosphodiesterase (PDE 2) has a low affinity catalytic domain and a cGMP specific allosteric domain. The low affinity catalytic site can hydrolyze both cAMP and cGMP with an apparent KM value lower for cGMP than for cAMP. However, when cGMP binds to the allosteric site, the catalytic site undergoes a change in conformation that shows high affinity for cAMP. PDE 2 shows the highest expression in the brain, but it is found in many other tissues as well and, therefore, has a wide potential functional and therapeutic utility (JA Beavo, et al., Rev. Physio. Biochem. Pharm. , 135 , 67 (1999)). Examples of the role and therapeutic potential of PDE 2 are in neuronal development, learning and memory (WCG van Staveren, et al., Brain Res., 888 , 275 (2001) and J. O'Donnell, et al. ., J. Pharm. Exp. Ther., 302 , 249 (2002)); secretion of prolactin and aldosterone (MO Velardez, et al., Eur. J. Endo., 143 , 279 (2000) and N. Gallo-Payet, et al., Endo., 140 , 3594 (1999)); Bone cell differentiation, growth, and bone resorption (C. Allardt-Lamberg, et al., Biochem. Pharm., 59 , 1133 (2000) and S. Wakabayashi, et al. J. Bone. Miner. Res., 17 , 249 (2002)); Immune response (MD Houslay, et al., Cell. Signal., 8 , 97 (1996); vascular angiogenesis (T. Keravis, et al., J. Vasc. Res., 37, 235 (2000); inflammatory cell transit (SL Wolda, et al., J. Histochem. Cytochem., 47 , 895 (1999); cardiac contraction (R. Fischmeister, et al., J. Clin. Invest., 99 , 2710 (1997), P. Donzeau -Gouge, et al., J. Physiol., 533 , 329 (2001), and DJ Paterson, et al., Card. Res., 52 , 446, (2001)); platelet aggregation (RJ Haslam, et al. ., Biochem. J., 323 , 371 (1997); female sexual arousal disorder (FSAD) (CP Wayman, et al., Publication of European patent application numbers EP 1 097 7707 and 1 0977 06); and vasoconstriction hypoxic pulmonary (J. Haynes, et al., J. Pharm. Exp. Ther., 276 , 752 (1996)). It has been shown that NASH (erythro-9- (2-hydroxy-3-nonyl) adenine) , a potent adenosine deaminase inhibitor, selectively inhibits PDE 2, however, the use of EHNA as a therapeutic agent based on PDE 2 est limited due to the low efficacy in the inhibition of PDE 2, and high efficacy in the inhibition of adenosine deaminase (R. Fischmeister, et al., Mol. Pharm., 48 , 121 (1995)).

It has now been found that certain derivatives of pyrido- [2,3-d] pyrimidin-2,4-diamine of the formula (I) later in this specification inhibit PDE 2 and, therefore, are useful in the treatment of disorders  physiological mediated by the cell signaling path of cAMP or cGMP.

United States patents numbers 5,547,954 and 5,710,157 describe certain 2,4-diamino-5,6-disubstituted- Y 5,6,7-trisubstituted-5-desazapteridines, the compositions thereof, and the uses thereof in the Insect control in agricultural crops.

WO 02/102 315 describes PDE7 inhibitors.

Summary of the invention

The invention provides compounds of formula (I)

one

and pharmaceutically salts acceptable compounds, in which n, X, and Y are as defined in this specification below; The compositions pharmaceuticals thereof; the combinations thereof; and the uses of same.

Detailed description of the invention

The present invention provides compounds of formula (I)

2

and the salts of the compounds pharmaceutically acceptable, in the that:

R 1 and R 2 are hydrogen or methoxy, with the condition that R1 and R2 are not both hydrogen or both methoxy;

n is 1, 2, 3, or 4;

X is a link; OR; S; C = O; -N (R) -; where R is hydrogen or (C 1 -C 3) alkyl; -C (OH) -; or -SO2; and

Y is benzoxazolyl; benzothiazolyl; benzofurazanyl; benzofuranyl; benzothiadiazolyl; bencisoxazolyl; bencisothiazolyl; benzimidazolyl; pyridyl; isatinyl; oxindolyl; indazolyl; indolyl; phenyl; thienyl; or furanyl; where and is optionally independently substituted with between one and three halogen; trifluoromethyl; methoxy; -C (= O) CH 3; cyano; -C (CH 3) 2 OH; -CH (CH 3) OH; -CH (CF 3) OH; -C (C = O) CF 3; -SO2 NH2; -C (= O) OCH 3; -CH 2 COOH;

3

thiazolyl; or oxadiazolyl.

A generally preferred subgroup of compounds of formula (I) comprises the compounds in which X is an E bond is benzofurazanyl; thienyl; pyridyl; or phenyl, in the which phenyl is optionally substituted independently with one or two halogen; trifluoromethyl; methoxy; -C (= O) CH 3; cyano; -C (CH 3) 2 OH; -CH (CH 3) OH; -CH (CF 3) OH; -C (C = O) CF 3; -SO2 NH2; -C (= O) OCH 3; -CH 2 COOH; thiazolyl; or oxadiazolyl.

A particularly preferred subgroup of compounds of formula (I) comprises the compounds in which X is a bond, n is 2 or 3, and Y is thienyl; pyridyl; or phenyl where phenyl is independently substituted with one or two methoxy; halogen; -C (CH 3) 2 OH; -CH (CF 3) OH or -C (C = O) CF 3.

A cyclic group can be linked to another group In more ways than one. If no provision of particular union, then all provisions are intended possible. For example, the term "pyridyl" includes 2-, 3-, or 4-pyridyl, and the term "thienyl" includes 2- or 3-thienyl

The compounds and intermediates of the present invention can be named according to the systems of Well nomenclature of the IUPAC (International Union of Pure Chemistry and Applied) or CAS (Chemical Abstracts Service, Columbus, OH).

The carbon atom content of various residues containing hydrocarbons can be indicated by a prefix that designates the minimum and maximum number of atoms carbon in the rest, that is, the prefix "alkyl (C_ {a} -C_ {b}) "indicates an alkyl moiety of whole number "a" a "b" of carbon atoms, inclusive.

The term "alkyl" means chains linear or branched, monovalent carbon atoms. The Examples of alkyl groups include methyl, ethyl, propyl, Isopropyl, butyl, isobutyl, and the like.

The term "halogen" represents chlorine, Fluoro, bromine, and iodine.

The term "mammal" means animals including, for example, dogs, cats, cows, sheep, horses, and Humans. Preferred mammals include humans from Any gender.

The term "salts" refers to the organic and inorganic salts of a compound of formula (I), or a prodrug thereof. These salts can be prepared in situ during the final isolation and purification of a compound, or by reacting separately a compound of formula (I), or a prodrug thereof, with a suitable organic or inorganic acid or base and salt isolation. so formed. Representative salts include the salts hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, besylate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, and lauryl sulphonate, and the like. These may also include cations based on alkali and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium, and the like, as well as non-toxic ammonium, quaternary ammonium and amine cations including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. For additional examples see for example, Berge et al., J. Pharm. Sci., 66 , 1-19 (1977).

The term "substituted" means that a hydrogen atom of a molecule has been replaced with an atom or different molecule. The atom or molecule that replaces the atom of Hydrogen is called a "substituent."

As used in this specification, the expression "therapeutically effective amount" means a amount of a compound that is capable of treating a condition Pathological described.

The terms "treat", "treatment", and "treating" include preventive treatment (for example, prophylactic) and palliative (for example, healing or healing), or the act of providing preventive or palliative treatment.

The compounds of formula (I) may contain asymmetric or chiral centers and, therefore, exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds and prodrugs of formula (I) as well as mixtures thereof, including racemic mixtures, form part of the present invention. In addition, the present invention encompasses all geometric and positional isomers. For example, if a compound or prodrug of formula (I) incorporates a double bond (s), both cis - and trans - forms, as well as mixtures thereof, are encompassed within the scope of the invention.

Diastereoisomeric mixtures can be separate into their individual diastereoisomers based on their physical-chemical differences by well known procedures by those ordinarily skilled in the art, such as by chromatography and / or fractional crystallization. The enantiomers are they can separate by converting the enantiomeric mixture into a mixture diastereomeric by reaction with an appropriate compound optically active (eg alcohol), separating the diastereomers and converting (for example, hydrolyzing) the individual diastereomers in pure enantiomers corresponding.

Compounds and prodrugs of compounds of formula (I) may exist in non-solvated forms as well as solvated with pharmaceutically acceptable solvents, such as water, ethanol, and the like, and the invention is intended to encompass both solvated and non-solvated forms.

It is also possible that the compounds and prodrugs of formula (I) may exist in the form of isomers tautomers in equilibrium, and all these forms are encompassed within the scope of the invention.

The present invention also encompasses compounds labeled with isotopes of formula (I), which are identical to those described in this specification, except for the fact that one or more atoms are replaced with an atom that has a mass atomic or mass number different from the atomic mass or mass number Usually found in nature. The examples of Isotopes that can be incorporated into the compounds of formula (I) include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F, and 36 Cl, respectively. The compounds of formula (I), the prodrugs thereof, and salts thereof pharmaceutically acceptable compounds and prodrugs, which contain the aforementioned isotopes and / or other isotopes of the other atoms are intended to be within the scope of the present invention

Certain isotope-labeled compounds of formula (I), for example, compounds in which radioactive isotopes such as 3 H and 14 C are incorporated, are useful in compound and / or distribution distribution assays. substrate fabrics. Tritiated isotopes, that is, 3 H, and with 14 C are particularly preferred for their relative ease of preparation and easy detection. In addition, substitution with heavy isotopes such as deuterium, i.e.2H, can produce certain therapeutic advantages that are produced by greater metabolic stability, for example, increased half-life in vivo , or reduced dosage requirements. Therefore, it may be preferred in some circumstances. Compounds of formula (I) labeled with isotopes can in general be prepared by performing procedures analogous to those described in the schemes and / or examples set forth herein below, substituting an unlabeled reagent with isotopes for an isotope labeled reagent. .

In another aspect, the invention provides the use of a compound of the formula (I) in the manufacture of a medication to treat conditions, diseases, or symptoms mediated by PDE 2 in a mammal in need of such treatment said procedure comprises administration to mammal of a therapeutically effective amount of a compound of formula (I), a prodrug thereof, or a salt of the compound or pharmaceutically acceptable prodrug; or a composition Pharmaceutical comprising a compound of formula (I), a prodrug thereof, or a salt of the compound or prodrug pharmaceutically acceptable, and a carrier, vehicle, or diluent pharmaceutically acceptable.

Conditions, diseases, or symptoms preferred that can be treated according to the present procedures include osteoporosis, pulmonary hypertension, disorder of female sexual arousal, memory or decreased cognition, platelet aggregation, vascular angiogenesis, dementia, cancer, arrhythmia, thrombosis, fracture and / or bone defect, delayed fracture or pseudoarthrosis, spinal fusion, bone growth inward, cranial facial reconstruction, or hypoxia. A Especially preferred condition is fracture and / or bone defect.

In another aspect, the invention provides the use of a compound of the formula (I) in the manufacture of a medicine to inhibit the activity of PDE 2 in a mammal in need for such inhibition said procedures comprise the administration to the mammal of a PDE 2 inhibitory amount of a compound of formula (I), or a salt of the compound pharmaceutically acceptable; or a pharmaceutical composition that comprises a compound of formula (I), or a salt of the compound pharmaceutically acceptable, and a carrier, vehicle, or diluent pharmaceutically acceptable.

The compounds of formula (I), and salts Pharmaceutically acceptable compounds, can be administer to a mammal at dosage levels in the range between about 0.001 mg and about 200 mg per day. For a normal human adult who has a body mass of approximately 70 kg, a dosage in typically the range between about 0.01 mg and about 100 mg per kg of body mass, however, some may be required variability in the general dosage range depending on the age and mass of the subject being treated, the proposed route of administration, the particular compound being administered, and the like The determination of the dosage intervals and optimal dosages for a particular mammalian subject is within the capacity of people of ordinary experience in the technique that have the benefit of the present description.

Still in another aspect, the invention provides pharmaceutical compositions comprising a combination of a PDE 2 inhibitor, according to the formula (I) a selective agonist of EP2; and a carrier, vehicle, or pharmaceutically acceptable diluent; and the procedures of treatment of osteoporosis, pulmonary hypertension, disorder of female sexual arousal, memory or decreased cognition, platelet aggregation, vascular angiogenesis, dementia, cancer, arrhythmia, thrombosis, fracture and / or bone defect, delayed fracture or pseudoarthrosis, spinal fusion, bone growth inward, cranial facial reconstruction, or hypoxia using such compositions. An especially preferred condition is fracture and / or bone defect.

Still in another aspect, the invention provides the use of a compound of the formula (I) in the manufacture of a medicine to treat fracture and / or bone defect in a mammal in need of such treatment said procedures comprise administration to the mammal of a therapeutically effective amount of a PDE 2 inhibitor, or a salt of said pharmaceutically acceptable inhibitor.

Any PDE 2 inhibitor, which includes the compounds of formula (I) in this specification, it can be used in the methods and combinations of the invention. The Examples of known PDE 2 inhibitors comprise EHNA, 6- (3,4-dimethoxybenzyl) -1 - ([(1-hydroxyethyl) -4-phenylbutyl] -3-methyl-1,5-dihydropyrazolo [3,4-d] pyrimidin-4-one  (BAY-60-7550; United States patent United No. 6,174,884), and 9- (1-acetyl-4-phenylbutyl) -2- (3,4-dimethoxybenzyl) -1,9-dihydropurin-6-one  (U.S. Patent No. 5,861,396). Additional examples of the PDE 2 inhibitors are described in the patents of United States numbers 5,861,396; 5,401,774; 6,458,796; and 6,555,547; and in the publication of the international application PCT Nº 98/32755.

Any selective receptor agonist EP2 can be used in the combination aspects of the present invention, however, a generally preferred class of the agonists of the selective receptor EP 2, described in the United States common assignment patent with this No. 6,498,172, comprising the compounds of formula AA

4

their prodrugs, and the pharmaceutically acceptable salts thereof, in which G, A, B, K, M, Q, and Z are as defined in that document.

Generally preferred compounds of formula AA are (3 - (((pyridin-3-sulfonyl) - (4-pyrimidin-5-ylbenzyl) amino) methyl) phenyl) acetic acid; (3 - (((5-phenylfuran-2-ylmethyl) - (pyridin-3-sulfonyl) amino) methyl) phenyl) acetic acid; (3 - (((pyridin-3-sulfonyl) - (4-pyrimidin-2-ylbenzyl) amino) methyl) phenyl) acetic acid; (3 - (((pyridin-3-sulfonyl) - (4-thiazol-2-ylbenzyl) amino) methyl) phenyl) acetic acid; (3 - (((4-pyrazin-2-ylbenzyl) - (pyridin-3-sulfonylamino) methyl) phenyl) acetic acid; (3 - (((4-cyclohexylbenzyl) - (pyridin-3-sulfonyl) amino) methyl) phenoxy) acetic acid; (3 - (((pyridin-3-sulfonyl) - (4-pyridin-2-ylbenzyl) amino) methyl) phenoxy) acetic acid; (3 - ((((pyridin-3-sulfonyl)) - (4-pyridin-3-ylbenzyl) amino) methyl) phenoxy) acetic acid (3 - (((pyridin-3-sulfonyl) - (4-pyridin-4-ylbenzyl) amino) methyl) phenoxy) acetic acid; (3 - (((pyridin-3-sulfonyl) - (4-thiazol-2-ylbenzyl) amino) methyl) phenoxy) acetic acid; 5- (3 - (((pyridin-3-sulfonyl) - (4-thiazole -2-ylbenzyl) amino) propyl) thiophene-2-carboxylic acid (3 - (((2,3-dihydrobenzo [1,4] dioxin-6-ylmethyl) - (pyridin-3-sulfonyl) amino) methyl) phenyl) acetic acid; and (3 - ((benzofuran-2-ylmethyl- (pyridin-3-sulfonyl) amino) methyl) phenyl) acetic acid; (3 - ((((4-butylphenyl) - (pyridine-3-sulfonyl) ) amino) methyl) phenyl) acetic acid; (3 - ((benzenesulfonyl) - (4-butylbenzyl) amino) methyl) phenyl) acetic acid; (3 - (((4-bu-
tilphenyl) - (1-methyl-1H-imidazol-4-sulfonyl) amino) methyl) phenyl) acetic acid; and acid (3 - (((4-dimethylaminobenzyl) - (pyridin-3-
sulfonyl) amino) methyl) phenyl) acetic acid; (3 - (((4-dimethylaminobenzyl) - (pyridin-3-sulfonyl) amino) methyl) phenoxy) acetic acid; and (3 - (((4- tert -butylbenzyl) - (pyridin-3-sulfonyl) amino) methyl) phenoxy) acetic acid; trans - (3 - (((3- (3,5-dichlorophenyl) allyl) - (pyridin-3-sulfonyl) amino) methyl) phenyl) acetic acid; (3 - (((2- (3,5-dichlorophenoxy) ethyl) - (pyridin-3-sulfonyl) amino) methyl) phenoxy) acetic acid; the prodrugs thereof, and the salts of the compounds and pharmaceutically acceptable prodrugs.

An especially preferred compound of formula AA is (3 - (((4- tert -butylbenzyl) - (pyridin-3-sulfonyl) amino) methyl) phenoxy) acetic acid, a prodrug thereof or a pharmaceutically prodrug salt or compound. acceptable. A particularly preferred salt is sodium salt.

Another generally preferred class of E2 selective receptor agonists useful in aspects of combination of the invention comprises the compounds, prodrugs, and pharmaceutically acceptable salts of formula BB plus below, which are described in the United States cession patent Common with this No. 6,288,120.

5

in which A, B, K, M, Q, and Z are as defined in that document.

Generally preferred compounds of formula BB are 7 - [(2'-hydroxymethylbiphenyl-4-ylmethyl) methanesulfonylamino] heptanoic acid; 7 - {[4- (3-hydroxymethylthiophene-2-yl) benzyl] methanesulfonylamino} heptanoic acid; 7 - [(2'-chlorobiphenyl-4-ylmethyl) methanesulfonylamino] heptanoic acid; 7 - {[4- (1-hydroxyhexyl) benzyl] methanesulfonylamino] heptanoic acid; 7 - [(4-Butylbenzyl) methanesulfonylamino] heptanoic acid; 7 - {[5- (1-hydroxyhexyl) thiophene-2-ylmethyl] methanesulfonylamino} heptanoic acid; (3 - {[(4-Butylbenzyl) methanesulfonylamino] methyl} phenyl) acetic acid; 7 - {[3- (3-chlorophenyl) propyl] methanesulfonylamino} heptanoic acid; 7 - {[3- (3,5-dichlorophenyl) propyl] methanesulfonylamino} hepta- acid
noico; 5- (3 - {[3- (3-chlorophenyl) propyl] methanesulfonylamino} propyl) thiophene-2-carboxylic acid; 7 - {[2- (3,5-dichlorophenoxy) ethyl] methanesulfonylamino} heptanoic acid; 5- (3 - {[2- (3,5-dichlorophenoxy) ethyl] methanesulfonylamino} propyl) thiophene-2-carboxylic acid; N- [2- (3,5-dichlorophenoxy) ethyl] -N- [6- (1H-tetrazol-5-yl) hexyl] methanesulfonamide; trans - (4 - {[3- (3,5-dichlorophenyl) allyl] methanesulfonylamino} butoxy) acetic acid; trans -N- [3- (3,5-dichlorophenyl) allyl] -N- [6- (1H-tetrazolyl-5-yl) hexyl] methanesulfonamide; trans - (5- (3 - {[3- (3,5-dichlorophenyl) allyl) methanesulfonylamino} propyl) thiophene-2-carboxylic acid; trans - [3 - ({[3- (3,5-dichlorophenyl) allyl] methanesulfonylamino} methyl) phenyl] acetic, the prodrugs thereof, and the salts of the pharmaceutically acceptable compounds and prodrugs.

An especially preferred compound of formula BB is acidic 7 - [(4-butylbenzyl) methanesulfonylamino] heptanoic acid,  a prodrug thereof, or a salt of the compound or prodrug pharmaceutically acceptable. A preferred salt is salt. monosodium

Other selective EP 2 receptor agonists useful in the combination aspects of the present invention comprise the prostaglandin receptor agonists described in US Patent Nos. 6,531,485; 6,376,533; 6,214,314; 5,877,211; 5,716,835; 5,698,598; and 5,462,968; U.S. Published Patent Application No. 2002/187961; N. Duckworth, et al Journal of Endocrinology, 172 (2), 263-269 (2002); K. Tani, et al., Synlett 2 , 239-242 (2002); K. Tani, et al., Bioorganic & Medicinal Chemistry, 10 (4), 1107-1114 (2002); K. Tani, et al., Bioorganic & Medicinal Chemistry, 10 (4), 1093-1106 (2002); J. Michelet, et al., EP 1 175 891 A1; K. Tani, et al., Bioorganic & Medicinal Chemistry Letters, 11 (15), 2025-2028 (2001); JY Crider, et al., International Journal of Environmental Studies, 58 (1), 35-46 (2000); JY Crider, et al., Journal of Ocular Pharmacology and Therapeutics, 17 , (1), 35-46 (2001); DF Woodward, et al., Journal of Ocular Pharmacology and Therapeutics, 11 , (3), 447-54 (1995); AT Nials, et al., Cardiovascular Drug Reviews, 11 (2), 165-79 (1993); and DF Woodward, et al., Prostaglandins, 46 (4), 371-83 (1993).

In the combination aspects of the invention, EP2 selective receptor agonists can be administered to mammals at dosage levels that vary between approximately 0.001 mg / kg and approximately 100 mg / kg mass body per day. For a normal adult human being of a mass body weight of approximately 70 kg, typically a dosage in the range of about 0.01 mg / kg and approximately 50 mg / kg of body mass, however, you can require some variability in the dosage range general depending on the age and mass of the subject being trying, the proposed route of administration, the compound particular that is being administered, and the like. the determination of the combination dosing intervals and dosages optimal for a particular mammalian subject is within the ability of those skilled in the art who have the benefit of This description.

Pharmaceutical compositions suitable for parenteral injection may comprise solutions, dispersions, suspensions, or emulsions, sterile aqueous and non-aqueous, and pharmaceutically acceptable sterile powders, for reconstitution extemporaneous in sterile injectable solutions or dispersions. The examples of vehicles, carriers, and aqueous diluents and not Suitable aqueous include water, ethanol, polyols (such as propylene glycol, polyethylene glycol, glycerol, and the like), suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters such as oleate of ethyl. You can maintain proper fluency, for example, by using a coating such as lecithin, by maintenance of the required particle size in the case of dispersions, and through the use of surfactants.

The pharmaceutical compositions of the invention may also comprise adjuvants, such as agents preservatives, humectants, emulsifiers, and dispersants. The prevention of contamination by microorganisms of the present compositions can be carried out with various agents antibacterials and antifungals, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. Can also be desirable to include isotonic agents, for example, sugars, chloride sodium, and the like. Prolonged absorption of the compositions Injectable pharmaceuticals can be done by using agents capable of delaying absorption, for example, monostearate  of aluminum and jelly.

Solid dosage forms for Oral administration include capsules, tablets, powders, and granules In such solid dosage forms, the compound active is mixed with at least one excipient (or vehicle) conventional inert pharmaceutical such as sodium citrate or phosphate dicalcium, or (a) fillers or extenders, such as starches, lactose, sucrose, mannitol, and silicic acid; (b) binders, such as for example, carboxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerol; (d) disintegrating agents, as per example, agar-agar, calcium carbonate, starch potato or tapioca, complex silicates of certain acids alginics, and sodium carbonate; (e) solution retarders, such as for example, paraffin; (f) absorption accelerators, as per example, quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h)  adsorbents, such as kaolin and bentonite; and / or (i) lubricants, such as talcum, calcium stearate, magnesium stearate, solid polyethylene glycols, lauryl sulfate sodium, or mixtures thereof. In the case of capsules and tablets, the dosage forms may further comprise buffering agents.

Solid compositions of a similar type are they can also be used as fillers in hard gelatin capsules and soft using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols, and the like.

Solid dosage forms such as tablets, dragees, capsules, and granules, can be prepared with coatings such as shells, such as coatings enteric and others well known to those skilled in the art. They can also comprise opacity agents, and they can also be of compositions such that they release the active compound (s) in a delayed manner, maintained, or controlled. Examples of inclusion compositions that can be used are polymeric substances and waxes. The active compound (s) can also be in microencapsulated form, if appropriate, with one or more of the excipients mentioned above.

Liquid dosage forms for Oral administration include emulsions, solutions, suspensions, Pharmaceutically acceptable syrups and elixirs. Besides active compounds, the liquid dosage form may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, as per example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, in particular, seed oil cotton, peanut oil, corn germ oil, oil Olive, castor oil, and sesame seed oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and esters of sorbitan fatty acids, or mixtures of these substances, and Similar.

In addition to such inert diluents, the Pharmaceutical composition may also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.

The suspensions, in addition to the active compound (s), may additionally comprise suspending agents, such as alcohols ethoxylated isostearyl, polyoxyethylene sorbitol esters and sorbitan, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, or mixtures of  the aforementioned substances, and the like.

Compositions for rectal administration or vaginal preferably comprise suppositories, which can be prepare by mixing a compound (s) asset (s) with suitable excipients or non-irritating vehicles such as cocoa butter, polyethylene glycol or a wax of suppositories that are ordinarily solid at room temperature, but fluids at body temperature, and therefore, melt in the rectum or vaginal cavity thereby releasing the component active.

Dosage forms for administration topical may include ointments, powders, sprays and inhalations The active compound (s) are Mix (n) in sterile condition with a vehicle, carrier, or pharmaceutically acceptable diluent, and any of the preservatives, buffers, or propellants that may be required.

A generally preferred composition for administer PDE 2 inhibitors, which include the compounds  of formula (I), as well as the combinations comprising the PDE 2 inhibitors and selective receptor agonists EP2 in the treatment of bone fractures, comprises a injectable, flowable composition that provides release held at a local injection site by forming a biodegradable solid or deposit of gel, matrix, or implant. A example of such an administration system comprises a system of distribution based on slow release polymers, biodegradable. See for example, the published patent application of the United States No. 2003-0104031 A1.

Such a polymer-based distribution system generally comprises a therapeutically active agent (s).
useful (s), dissolved (s) or dispersed (s) in a flowable, biodegradable, thermoplastic polymer solution or dispersion in an organic solvent. Upon injection of the composition, the organic solvent diffuses out of the injection site, causing the polymer to precipitate or gel, thereby trapping the agent (s) in a sustained release reservoir. The agent (s) are subsequently released by diffusion, and erosion, of the polymer matrix. The matrix is slowly eroded by hydrolysis and eventually disappears from the site of administration. The molecular weight and concentration of the polymer can control the release in vivo of the agent (s) as well as the rate of degradation of the matrix.

The polymer-based distribution system provides sustained release of an active agent (s) in vivo for a sustained period of time with minimal or reduced momentum in a patient in need thereof. A large discharge of agent (s) would result in poor local tolerance due to local effects (eg irritation) and minimize the amount of agent (s) available for effectiveness. The advantage that this administration procedure offers is that it minimizes or reduces the initial boost, but still releases the agent (s) to effective levels for sustained periods of time after a single local injection.

The polymer system is prepared by putting in contact the flowable composition with a gelation medium to coagulate or gel the composition in a solid, polymeric matrix microporous, or a polymeric gel matrix. The flowable composition contains a thermoplastic polymer or copolymer together with a suitable solvent. The polymers or copolymers, which form the matrix body, are substantially insoluble, so preferable essential and completely insoluble, in water and fluids bodily The insolubility of the matrix body allows it to function as an individual site for controlled release of (of the) agent (s). Polymers or copolymers are also biocompatible and biodegradable and / or bioerodible within the body of an animal, for example, mammal. Biodegradation allows the patient metabolizes and excretes the polymer matrix so that There is no need for surgical removal. Because the flowable composition and the polymeric system are biocompatible, the insertion procedure and the presence of the polymer system inside the body they do not produce substantial tissue irritation or Necrosis in the implant system. The composition of this invention is administered in the form of a flowable composition directly in body tissues.

Thermoplastic polymers suitable for incorporation into the solid matrix of the release system controlled are solid, pharmaceutically compatible and biodegradable by cellular action and / or by the action of body fluids. Examples of thermoplastic polymers suitable include polyesters of diols and carboxylic acids or of hydroxycarboxylic acids, such as polylactides, polyglycolides, and the copolymers thereof. Plus preferably the polymer comprises the copolymer, poly (acid lactic-coglycolic) (abbreviated PLGH) that, After hydrolysis, it produces lactic and glycolic acid. Downloading release of this copolymer can also be minimized by addition of polyethylene glycol (PEG) to form the PLGH blocked in the end with PEG.

Preferred materials comprise polylactides, polyglycolides, and copolymers thereof. These  polymers can be used to take advantage of the system polymer partly because they show excellent biocompatibility They cause little, if any, tissue irritation, inflammation, necrosis, or toxicity. In the presence of water, the polymers produce lactic acid and glycolic acid, respectively, They are easily metabolized. Polylactides can also incorporate glycolide monomer to enhance degradation resulting polymeric. These polymers are also preferred due to that effectively control the release rate of the agent (s) of the polymer system, and why they result local retention of the agent (s) at the site of administration.

The solubility or miscibility of a polymer thermoplastic in the organic solvent of the composition will vary according to factors such as crystallinity, hydrophilicity, capacity of binding to hydrogen, and the molecular weight of the polymer. By consequently, the molecular weight and the concentration of the polymer in the solvent is adjusted to achieve the desired miscibility as well as a desired release rate for the agent (s) incorporated.

The flowable polymer composition thermoplastic, solvent and the agent (s) It comprises a stable flowable substance. Preferably occurs a homogeneous solution of the agent (s) in a organic solvent The thermoplastic polymer is substantially soluble in organic solvent. After placement of the fluid composition in the body, the solvent dissipates and the polymer solidifies or gels to form the polymer system that has the agent (s) within an array solid polymer or gel.

For certain preferred polymers, the weight molecular polymer or copolymer is adjusted to be inside from a range of about 0.2 to about 0.4 of inherent viscosity (V. I. in deciliters / g) for release Sustained effective of bone growth promoter compound. The typical release rate of agent (s) Built-in (s) occurs at a V. I. of approximately 0.2 (approximately 8,000 to approximately 16,000 molecular weight) or about 0.3 (about 23,000 to about 45,000 molecular weight), but may vary depending on the particular components of the composition. For most of the systems, it is preferred to adjust the molecular weight of the polymer to approximately 0.2 V. V. for an effective release of (from the agents).

For a poly (DL-lactide) or a polymeric system of lactide-co-glycolide, the interval desired molecular weight is between about 0.2 and approximately 0.4 of V. I., with a value of V. I. of about 0.2. The molecular weight of a polymer can be Modify by conventional procedures.

Commercially thermoplastic polymers available, especially preferred include the following: PLGH copolymer with a 1: 1 ratio of lactic and glycolic acid with an inherent viscosity of approximately 0.2 dl / g (commercially available from Boehringer Ingelheim as a copolymer RESOMER® RG 502 H) (approximately 12,000 molecular weight); he PLGH copolymer with a 1: 1 ratio of lactic and glycolic acid with an inherent viscosity of approximately 0.3 dl / g (commercially available from Boehringer Ingelheim as a copolymer RESOMER® RG 503 H) (approximately 37,000 molecular weight); he PLGH copolymer with a 1: 1 embodiment of lactic acid and glycolic with an inherent viscosity of about 0.4 dl / g (commercially available from Boehringer Ingelheim as a copolymer RESOMER® RG 504 H) (approximately 47,000 molecular weight); Y PLGH copolymer with polyethylene glycol (PEG) terminal cap with a 1: 1 ratio of lactic and glycolic acid with a inherent viscosity of approximately 0.79 dl / g (commercially available from Boehringer Ingelheim as PLG-PEG) (approximately 52,000 molecular weight).

The solvents used in the thermoplastic compositions are preferably pharmaceutically acceptable, biocompatible, and will dissipate in the body fluid in situ so that they can be classified as having a water solubility that varies between highly soluble and insoluble. Preferably, they produce relatively little, if any, tissue irritation or necrosis at the site of injection and implantation. Preferably, the solvent will have at least a minimum degree of water solubility. When the organic solvent is water soluble or minimally soluble in water, the solvent will slowly disperse with the flowable polymer composition. The result will be an implant, which during the course of its life, may contain varying amounts of residual solvent. Preferably, the organic solvent has a moderate to high degree of water solubility so that it will readily disperse with the polymer composition in body fluids. Most preferably, the solvent is rapidly dispersed with the polymer composition so that a solid implant is rapidly formed. Simultaneous with the dispersion of the solvent, the thermoplastic polymer coagulates or gels in the solid polymer system. Preferably, as the thermoplastic polymer coagulates, solvent dispersion produces pore formation in the polymer system. As a result, the flowable composition containing the thermoplastic polymer, solvent, and agent (s) will form a porous solid polymer system. Also, when the solvent is slightly soluble in water, or is insoluble in water, solvent dispersion may result in the formation of a porous solid implant, or if some solvent remains with the implant, the result may be the formation of a gel implant that has few or no pores.

Suitable solvents include the liquid organic compounds that meet the above criteria. A generally preferred solvent comprises N-methyl-2-pyrrolidone (NMP)

Solvents for flowable compositions thermoplastic polymers are chosen for compatibility and Appropriate solubility of the polymer and solvent. Polymers Thermoplastics of lower molecular weight will normally dissolve more easily in solvents than high weight polymers molecular. As a result, the polymer concentration thermoplastic dissolved in the various solvents differs depending on the type of polymer and its molecular weight. In contrast, thermoplastic polymers of molecular weight more high will tend to coagulate, gel or solidify more quickly than thermoplastic molecular weight polymers very low. In addition, low molecular weight polymers tend to provide higher solution viscosities than materials Low molecular weight. So, for an injection efficiency advantageous, in addition to the advantageous release rate, it control the molecular weight and concentration of the polymer in the solvent

After formation of the polymer system from the flowable composition, the agent (s) is incorporated into the polymer matrix. After insertion of the flowable composition to form the polymer system, the agent (s)
te (s) is released from the matrix in adjacent tissues or fluids through diffusion and degradation mechanisms. The manipulation of these mechanisms can also influence the release of the agent (s) in the surroundings at a controlled rate. For example, the polymer matrix can be formulated to degrade after an effective and / or substantial amount of the agent (s) is released from the matrix. Thus, the release of the agency
te (s) of the matrix can be varied, for example, the solubility of the agent (s) in water, the distribution of the bone growth promoter compound with the matrix, or the size, shape, porosity, solubility, and biodegradability of the polymer matrix, among other factors. The release of the matrix agent (s) is controlled relative to its inherent speed by varying the polymer molecular weight to provide a desired duration and rate of release.

For example, a preferred dosage form of the agent (s) comprises a lyophilisate to reconstitute with a solution of PLGH in NMP before administration. The dosage form, constituted by the lyophilized compound in a syringe (syringe A) and a solution of PLGH in NMP in a second syringe (syringe B), is known as the A / B reconstitution system. The contents of both syringes are mix immediately before distributing the dose in one place or near him. After reconstitution, the contents are transferred to a graduated dosing syringe for administration. The dosage forms administered will be a solution and will result in the dispersion of the compound with PLGH in NMP at the desired potencies of, for example, 5 and 50 mgA / ml (mgA / ml refers to the free acid equivalent of the salt form sodium of the agent (s). The dosage form is a parenteral sustained release injection (for example, subcutaneous, intramuscular, or intramedullary) for administration local. This compound in a slow-release polymer matrix (prolonged release injection) is designed for administration on or near a site, and is not intended for intravenous administration To provide stability Adequate shelf life for dosage form, can be used a two syringe system (A / B), as described above,  preferably with the sodium salt form of the compound. A uniform formulation, preferably in the form of free acid of the compound, is a preferred alternative formulation. Based in the stability of the agent (s) and of the polymer, may prefer filtration of the agent (s) and irradiation of the polymer solution for the manufacture of a stable sterile product. In one embodiment, the form of Dosage can be manufactured and transported in the form of bags of separate aluminum containing syringes filled with the shape lyophilized agent (s) in a bag and solution polymeric in the other bag. The vessels, systems, and distribution procedures for lyophilization of bone growth promoter compounds are described in the PCT international patent application publication published No. WO 01/73363. Other administration procedures include local administration by injection to a particular site or distribution through a catheter to a site. The examples Additional information can be found in the provisional application for United States No. 60 / 335,156, filed on November 30, 2001

The teachings of all state patents United described in this specification are incorporated as reference in its entirety.

The compounds of formula (I), and salts Pharmaceutically acceptable compounds, can be prepared according to the exemplary synthetic pathways described in the schemes and examples in this specification below, as well as by other conventional organic preparatory procedures known, or obvious in the light of the present description, for experts in the relevant art. The procedures described in The present schemes are intended for purposes of exemplification of the present invention and should not be interpreted in no way as limitations thereof.

Scheme one

6

In scheme 1, stage 1, 2,4-dichloropyrido [2,3-d] pyrimidine (IV) is reacted with a benzylamine properly substituted (V) in the presence of a trisubstituted amine base, such such as triethylamine (TEA) or diisopropylethylamine (DIPEA), or a base aromatic, such as pyridine. The reaction is typically carried out in a polar alcoholic solvent, such as methanol (MeOH), ethanol (EtOH), or isopropanol (IPA), at a temperature that varies between about 0 ° C and about 100 ° C. Preferably, the reaction is carried out in the presence of DIPEA in ethanol at approximately room temperature (TA). In stage 2, the resulting condensation product (VI) is then reacted with an appropriately substituted amine (VII) in the presence of a trisubstituted amine base, such as TEA or DIPEA, or a base aromatic, such as pyridine, to produce compound (I). Typically the reaction is carried out in a polar aprotic solvent, such as N, N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), N-methylpyrrolidinone, or sulfolane, at an elevated temperature that varies between about 60 ° C and about 250 ° C. Preferably, the reaction is carried out in the presence of DIPEA in DMSO between about 90 ° C and about 120 ° C.

Although Scheme 1 has been shown as a two-step separate process in which intermediate (VI) is isolated and then reacted with amine (VII), it has also been found convenient to prepare and react (IV) in situ. with the amine (VII) in a single stage. In such a procedure, an aprotic solvent, preferably DMSO, is used. This reaction is also carried out in the presence of DIPEA in DMSO at a temperature between about 90 ° C and about 120 ° C.

Preparatory experimental section

Unless otherwise indicated, all Reagents used were obtained commercially. Unless I know indicate otherwise, the following experimental abbreviations They have the meanings indicated:

AcOH

- acetic acid

desc

- decomposition

DMPA

- 4-dimethylaminopyridine

EtOAc

- ethyl acetate

hr

- hours)

THE H

- lithium aluminum hydride

min

- minutes (s)

EM

- Mass spectrometry

NMR

- nuclear magnetic resonance

THF

- tetrahydrofuran

p -TsOH

- p -poluenesulfonic acid

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Preparation one

(2-Chloropyrido [2,3-] pyrimidin-4-yl) - (3,5-dimethoxybenzyl) amine

To a stirred solution of 2,4-dichloropyrido [2,3-d] pyrimidine (1.3 g, 6.7 mmol) and 3,5-dimethoxybenzylamine (1.1 g, 6.7 mmol) in 30 ml of EtOH at RT TEA (4 ml, 28.7 mmoles). A precipitate formed which was filtered off and washed with cold EtOH followed by hexanes to provide 1.8 g of title compound (82%) in the form of a solid. p. of f. 185 ° C (des). 1 H-NMR (DMSO-d 6) δ: 9.5 (t, 1H), 8.9 (dd, 1H), 8.7 (dd, 1H), 7.5 (m, 1H), 6.5 (d, 2H), 6.4 (t, 1H), 4.6 (d, 2H), 3.7 (s, 6H). MS (m / z,%): 331 (100)

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Preparation 2

(2-Chloropyrido [2,3-] pyrimidin-4-yl) - (3,4-dimethoxybenzyl) amine

This compound was prepared in an analogous manner. to that described in preparation 1 using the starting materials appropriate. 1 H-NMR (DMSO-d6) δ: 9.5 (t, 1H), 8.9 (dd, 1H), 8.7 (dd, 1H), 7.0 (s, 1H), 6.9 (d, 2H), 4.6 (d, 2H), 3.7 (s, 3H) 3.7 (s, 3H). MS (m / z,%): 331 (100).

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Preparation 3

2- (4-Aminomethylphenyl) propan-2ol

To a stirred solution of 4- (1-hydroxy-1-methylethyl) benzonitrile (2.0 g, 12.4 mmol) in 30 ml of THF at 0 ° C was added dropwise 1.0 N LAH in THF (26 ml, 26.1 mmol). The mixture was allowed to warm. until TA, and then heated at reflux for 20 minutes. The mixture was then cooled to 0 ° C and quenched with 5 ml of MeOH.  added drop by drop. The mixture was diluted with 300 ml of chloroform and washed with water (1 x 80 ml), dried over magnesium sulfate, and concentrated to provide 1.9 g (95%) of the title compound in the form of a solid. 1 H-NMR (CDCl 3) δ: 7.45 (d, 2H), 7.26 (d, 2H), 3.83 (s, 2H), 1.57 (s, 6H). GC-MS (m / e,%): 164 (M +, 15), 150 (80), 132 (75), 106 (100).

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Preparation 4

3- (4-Acetylphenyl) propionitrile

A mix of 4- (2-Bromoethyl) acetophenone (2.0 g, 8.8 mmol) and KCN (0.6 g, 8.8 mmol) in 30 ml of DMSO was heated to 75 ° C for 4 hours. The mixture was diluted with water and extracted with EtOAc The organic extract was washed successively with water and Brine, dried, and concentrated to provide an oil. The chromatography on silica gel eluting with 40% EtOAc / hexanes provided 0.8 g of an oil. 1 H-NMR (CD 3 OD) δ: 7.9 (d, 2H), 7.4 (d, 2H), 3.3 (t, 2H), 2.8 (t, 2H), 2.6 (s, 3H). GC-MS (m / e,%): 173 (M +, 20), 158 (100).

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Preparation 5

3- [4- (1-Hydroxy-1-methylethyl) phenyl] propionitrile

To a stirred solution of chloride 3M methylmagnesium in THF (3.3 ml, 9.8 mmol), diluted additionally with 10 ml of THF, a solution was added dropwise 3- (4-acetylphenyl) propionitrile (0.7 g, 3.9 mmol) in 10 ml of THF at -40 ° C. The reaction mixture was left. warm slowly to RT overnight, cooled to 0 ° C, after quenching with aqueous AcOH added dropwise. The reaction mixture was diluted with water and extracted with EtOAc. He organic extract was washed successively with water and brine, it was dried, and concentrated to provide 0.8 g of an oil. 1 H-NMR (CD 3 OD) δ: 7.4 (d, 2H), 7.2 (d, 2H), 2.9 (t, 2H), 2.7 (t, 2H), 1.5 (s, 6H). GC-MS (m / e,%): 189 (M +, 5), 174 (100).

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Preparation 6

2- [4- (3-Aminopropyl) phenyl] propan-2-ol

To a stirred solution of 1 M LAH in THF, diluted subsequently with 20 ml of THF, at 0 ° C was added dropwise to drop a solution of 3- (4- (1-hydroxy-1-methylethyl) phenyl] propionitrile (0.8 g, 4.0 mmol) in 10 ml of THF. The reaction was allowed to warm up. slowly until TA then heated to reflux for 4 hours. The reaction mixture was then cooled to 0 ° C and quenched with MeOH added drop by drop slowly. The mixture was diluted with chloroform and washed with water. The organic extract was filtered to through diatomaceous earth, the concentrated filtrate is then diluted with ethyl acetate, dried, and concentrated to provide 0.5 g of an oil. 1 H-NMR (CD 3 OD) δ: 7.4 (d, 2H), 7.1 (d, 2H), 2.6 (m, 4H), 1.7 (m, 2H), 1.5 (s, 6H). MS (m / e,%): 194 (M + +1, 100), 176 (90).

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Preparation 7

3- [4- (2-Methyl- [1,3] dioxolan-2-yl) phenyl] propionitrile

A mixture of 3- (4-acetylphenyl) propionitrile (2.2 g, 13 mmol), ethylene glycol (2.8 ml, 51 mmol), and a catalytic amount of p- TsOH (approximately 200 mg) in 100 ml of toluene He heated to reflux in a Dean Stark trap for 18 hours. The mixture was diluted with EtOAc and washed successively with 5% sodium bicarbonate solution, water and brine, dried (MgSO4), and concentrated to provide an oil. Chromatography on silica gel eluting with EtOAc / hexane solution provided 2.5 g of an oil. 1 H-NMR (CDCl 3) δ: 7.4 (d, 2H), 7.2 (d, 2H), 4.0 (m, 2H), 3.8 (m, 2H ), 2.9 (t, 2H), 2.6 (t, 2H), 1.6 (s, 3H). MS (m / e,%): 216 (M + - 1, 1), 202 (100).

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Preparation 8

3- [4- (2-Methyl [1,3] dioxolan-2-yl) phenyl] propylamine

To a stirred solution of 3- [4- (2-methyl- [1,3] dioxolan-2-yl) phenyl] propionitrile (2.4 g, 11 mmol) in 30 ml of THF was added dropwise 1 M LAH solution in THF. The mixture was allowed to warm to RT then heated at reflux for 1 hour. The mixture cooled until 0 ° C after quenching with MeOH added dropwise. The The mixture was diluted with chloroform and washed with water. The suspension resulting was filtered through diatomaceous earth and separated The filtered phases. The organic extract was dried over MgSO4 and concentrated to provide 2.4 g of an oil. 1 H-NMR (CDCl 3) δ: 7.4 (m, 2H), 7.1 (d, 2H), 4.0 (m, 2H), 3.8 (m, 2H), 2.7 (m, 2H), 2.6 (m, 2H), 1.7 (m, 2H), 1.6 (s, 3H). MS (m / e,%): 221 (M +, 10), 206 (60), 189 (100)

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Preparation 9

2,2,2-Trifluoro-1- (4-iodophenyl) ethanol

To a stirred solution of 4-iodobenzaldehyde (2.0 g, 8.6 mmol) in 20 ml of THF at RT a 0.5 M solution of trimethyl (trifluoromethyl) silane in THF (19 ml, 9.5 mmol) followed by tetrabutylammonium fluoride (112 mg, 0.4 mmoles). The mixture was stirred at RT overnight, poured in 0.1 N hydrochloric acid, and extracted with EtOAc. Turn the Organic extract was washed with water and brine, dried (MgSO4) and concentrated to provide 2.6 g of an oil. 1 H-NMR (CDCl 3) δ: 7.7 (d, 2H), 7.2 (d, 2H), 5.0 (m, 1H), 2.7 (d, 1H). MS (m / e,%): 302 (M +, 100), 233 (100).

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Preparation 10

2- {3- [4- (2,2,2-Trifluoro-1-hydroxyethyl) phenyl] prop-2-inyl} isoindole-1,3-dione

To an agitated suspension of 2,2,2-trifluoro-1- (4-iodophenyl) ethanol  (2.6 g, 8.5 mmol), N-propargylphthalimide (1.6 g, 8.5 mmol), dichlorobis (triphenylphosphine) palladium (298 mg, 0.43 mmol) and copper (I) iodide (82 mg, 0.43 mmol) in 20 ml of THF at RT 5 ml of TEA was added. The mixture was deaerated briefly in a stream of nitrogen and then heated to reflux for 6 hours. The mixture was diluted with chloroform, washed with water, dried over MgSO4, and concentrated to Provide a solid. The solid was triturated with EtOAc to produce 2.2 g of a solid. 1 H-NMR (CDCl 3) δ: 7.9 (m, 2H), 7.7 (m, 2H), 7.5 (d, 2H), 7.4 (d, 2H), 5.0 (m, 1H), 4.7 (s, 2H), MS (m / e,%): 359 (M +, 100).

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Preparation eleven

2- {3- [4- (2,2,2-Trifluoro-1-hydroxyethyl) phenyl] propyl} isoindole-1,3-dione

A mixture of 2- {3- [4- (2,2,2-trifluoro-1-hydroxyethyl) phenyl] prop-2-inyl} isoindole-1,3-dione (2.2 g, 6.1 mmol) and 10% Pd / C (220 mg) in 150 ml of EtOH and 150 ml of THF were stirred at 50 psi (344.83 kPa) of hydrogen at RT in a Parr apparatus for 2 hours. An additional 220 mg of 10% Pd / C was added and the mixture was stirred at 50 psi (344.83 kPa) of hydrogen at RT for an additional 2 hours. Then an additional 220 mg of 10% Pd / C was added and the mixture was stirred at 50 psi (344.83 kPa) of hydrogen at RT overnight. The mixture was filtered through diatomaceous earth and the filtrate was concentrated to provide an oil. Chromatography on silica gel eluting with EtOAc / hexanes gave 1.8 g of an oil. 1 H-NMR (CDCl 3) δ: 7.8 (m, 2H), 7.7 (m, 2H), 7.3 (d, 2H), 7.2 (d, 2H ), 4.9 (m, 1H), 3.7 (t, 2H), 2.7 (t, 2H), 2.0 (m, 2H). MS (m / e,%): 363 (M +, 15), 345 (35), 325 (50), 161
(100)

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Preparation 12

1- [4- (3-Aminopropyl) phenyl] 2,2,2-trifluoroethanol

To a stirred suspension of 2- {3- [4- (2,2,2-trifluoro-1-hydroxyethyl) phenyl] propyl} isoindole-1,3-dione (1.8 g, 5.0 mmol), in 50 ml of MeOH at RT hydrazine hydrate (0.46 ml, 15.0 mmol) was added. The mixture was stirred at RT for 18 hours. The reaction mixture was filtered and the filtrate was concentrated to provide an oil. The oil was triturated with chloroform, filtered, and the filtrate was concentrated to provide 1.0 g of an oil. 1 H-NMR (CDCl 3) δ: 7.3 (d, 2H), 7.2 (d, 2H), 5.0 (m, 1H), 2.6 (m, 2H ), 2.5 (m, 2H), 1.6 (m, 2H), MS (m / e,%): 363 (M +, 10), 216
(100)

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Preparation 13

Ester benzo [1,2,5] oxadiazol-5-yl trifluoromethanesulfonic acid

To a stirred solution of 5-hydroxybenzofurazan (1.8 g, 13.0 mmol), TEA (2.4 ml, 33.0 mmol) and DMAP (79 mg, 7.0 mmol) in 40 ml of dichloromethane at - 78 ° C a solution of trifluoromethanesulfonic anhydride (2.8 ml, 17.0 mmol) in 10 ml of dichloromethane was added dropwise. The mixture was allowed to slowly warm to RT for 3 hours, diluted with dichloromethane, and washed with water. The organic extract was dried and concentrated to provide an oil. Chromatography on silica gel eluting with dichloromethane produced 3.1 g of an oil. 1 H-NMR (CDCl 3) δ: 8.0 (dd, 1H), 7.8 (dd, 1H), 7.3 (dd, 1H). MS (m / e,%): 268 (M +, 60), 146 (60), 69
(100)

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Preparation 14

2- (Benzo [1,2,5] oxadiazol-5-ylprop-2-inyl) isoindole-1,3-dione

This compound was prepared in an analogous manner. to that of preparation 10 using appropriate starting materials. 1 H-NMR (CDCl 3) δ: 8.2 (s, 1H), 8.0 (dd, 1H), 7.9 (m, 2H), 7.8 (m, 2H), 7.5 (dd, 2H), 4.7 (s, 2, 2H). MS (m / e,%): 303 (M +, 100).

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Preparation fifteen

2- (3-Benzo [1,2,5] oxadiazol-5-ylpropyl) isoindole-1,3-dione

This compound was prepared in an analogous manner. to that of preparation 11 using appropriate starting materials. 1 H-NMR (CDCl 3) δ: 7.8 (m, 2H), 7.7 (m, 3H), 7.6 (s, 1H), 7.2 (m, 1H), 3.8 (t, 3H), 2.8 (t, 2H), 2.1 (m, 2H). MS (m / e,%): 307 (M +, 40), 290 (30), 272 (20), 160 (100)

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Preparation 16

3-Benzo [1,2,5] oxadiazol-5-ylpropylamine

This compound was prepared in an analogous manner. to that of preparation 12 using appropriate starting materials. 1 H-NMR (DMSO-d 6) δ: 7.9 (m, 2H), 7.7 (m, 1H), 7.5 (m, 1H), 2.7 (m, 2H), 2.5 (m, 2H), 1.7 (m, 2H). MS (m / e,%): 177 (M +, 40), 160 (100).

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Preparation 17

Benzothiazol-6-Ilic acid ester trifluoromethanesulfonic

This compound was prepared in an analogous manner. to that of preparation 13 using appropriate starting materials. 1 H-NMR (CDCl 3) δ: 9.1 (s, 1H), 8.2 (d, 1H), 7.9 (d, 1H), 7.4 (dd, 1H). MS (m / e,%): 283 (M +, 80), 150 (100)

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Preparation 18

2- (3-Benzothiazol-6-ylprop-2-inyl) isoindole-1,3-dione

This compound was prepared in an analogous manner. to that of preparation 10 replacing 2,2,2-trifluoro-1- (4-iodophenyl) ethanol  by the benzothiazol-6-yl acid ester trifluoromethanesulfonic acid (preparation 17). 1 H-NMR (DMSO-d 6) δ:  9.4 (s, 1H), 8.3 (d, 1H), 8.0 (d, 2H), 7.9 (m, 1H), 7.8 (m, 2H), 7.5 (dd, 1H), 4.6 (s, 2H). MS (m / e,%): 318 (M +, 100).

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Preparation 19

2- (3-Benzothiazol-6-ylpropyl) isoindole-1,3-dione

This compound was prepared in an analogous manner. to that of preparation 11 replacing 2- {3- [4- (2,2,2-Trifluoro-1-hydroxyethyl) phenyl] prop-2-inyl} isoindole-1,3-dione  (preparation 18) by the 2- (3-Benzothiazol-6-ylprop-2-inyl) isoindole-1,3-dione (preparation 10). 1 H-NMR (CDCl 3) δ: 8.9 (s, 1H), 8.0 (d, 1H), 8.0 (d, 1H), 7.8 (m, 2H), 7.7 (m, 2H), 7.3 (dd, 1H), 3.8 (t, 2H), 2.8 (t, 2H), 2.1 (m, 2H). EM (m / e,%): 332 (M +, 80), 174 (40), 162 (100).

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Preparation twenty

3-Benzothiazol-6-ylpropylamine

This compound was prepared in an analogous manner. to that of preparation 12 replacing the 2- {3- [4- (2,2,2-Trifluoro-1-hydroxyethyl) phenyl] propyl} isoindole-1,3-dione  (preparation 11) by the 2- (3-benzothiazol-6-ylpropyl) isoindole-1,3-dione (preparation 19). MS (m / e,%): 192 (M +, 20), 175 (100).

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Preparation twenty-one

2- (3-Iodophenyl) -2-methyl [1,3] dioxolane

A mixture of 3-iodoacetophenone (3.0 g, 12 mmol), ethylene glycol (2.7 ml, 48 mmol), and p- toluenesulfonic acid (30 mg) in 50 ml of toluene was heated to reflux in a Dean trap. Stark for 18 hours. The mixture was diluted with EtOAc and washed successively with 5% sodium bicarbonate solution, water, and brine, dried over MgSO4, and concentrated to provide an oil. Chromatography on silica gel eluting with EtOAc / hexanes gave 3.1 g of an oil. 1 H-NMR (CDCl 3) δ: 7.8 (m, 1H), 7.6 (dd, 1H), 7.4 (dd, 1H), 7.0 (m, 1H ), 4.0 (m, 2H), 3.7 (m, 2H), 1.6 (s, 3H). MS (m / e,%): 290 (M +, 20), 275 (100).

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Preparation 22

2- {3- [3- (2-Methyl [1,3] dioxolan-2-yl) phenyl] prop-2-inyl} isoindole-1,3-dione

This compound was prepared in an analogous manner. to that of preparation 10 replacing the 2,2,2-trifluoro-1- (4-iodophenyl) ethanol  by 2- (3-iodophenyl) -2-methyl [1,3] dioxolane (preparation 21). 1 H-NMR (CDCl 3) δ: 7.9 (m, 2H), 7.7 (m, 2H), 7.5 (s, 1H), 7.4 (m, 1H), 7.3 (m, 1H), 7.2 (m, 1H), 4.7 (s, 2H), 4.0 (m, 2H), 3.7 (m, 2H), 1.6 (s, 6H). EM (m / e,%): 347 (M +, 15), 332 (100).

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Preparation 2. 3

2- {3- [3- (2-Methyl [1,3] dioxolan-2-yl) phenyl] propyl} isoindole-1,3-dione

This compound was prepared in an analogous manner. to that of preparation 11 replacing the 2- {3- [4- (2,2,2-Trifluoro-1-hydroxyethyl) phenyl] prop-2-inyl} isoindole-1,3-dione (preparation 10) by 2- {3- [3- (2-methyl [1,3] dioxolan-2-yl) phenyl] prop-2-inyl} isoindole-1,3-dione (preparation 22). 1 H-NMR (CDCl 3) δ: 7.8 (m, 2H), 7.7 (m, 2H), 7.2 (m, 4H), 7.1 (d, 1H), 4.0 (m, 2H), 3.7 (m, 4H), 2.6 (t, 2H), 2.0 (m, 2H), 1.6 (s, 3H). EM (m / e,%): 351 (M +, 5), 336 (100).

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Preparation 24

3- [3- (2-Methyl [1,3] dioxolan-2-yl) phenyl] propylamine

This compound was prepared in an analogous manner. to that of preparation 12 replacing the 2- {3- [4- (2,2,2-Trifluoro-1-hydroxyethyl) phenyl] propyl} isoindole-1,3-dione  by 2- {3- [3- (2-methyl [1,3] dioxolan-2-yl) phenyl] propyl} isoindole-1,3-dione  (preparation 23). 1 H-NMR (CDCl 3) δ: 7.3 (m, 3H), 7.1 (dd, 1H), 4.0 (m, 2H), 3.8 (m, 2H), 2.7 (t, 2H), 2.6 (t, 2H), 1.8 (m, 2H), 1.6 (s, 3H). MS (m / e,%): 221 (M +, 20), 206 (60), 189 (100).

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Preparation 25

2- (3-Iodophenyl) propan-2-ol

To a stirred solution of chloride methylmagnesium (65 mmol) in 100 ml of THF at 0 ° C was added dropwise to 3-iodoacetophenone drop (4.0 g, 16.3 mmol). The reaction mixture was allowed to warm to RT then cooled to 0 ° C and an additional equivalent of chloride of methylmagnesium The mixture was allowed to warm to RT and stirred. for 5 hours The reaction mixture was quenched with MeOH, diluted with water, acidified with glacial AcOH and extracted with dichloromethane The organic extract was washed with solution of 5% sodium bicarbonate was then concentrated. Chromatography on silica gel eluting with dichloromethane provided a oil that solidified after rest. MS (m / e,%): 262 (M +, 80), 247 (100).

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Preparation 26

2- {3- [3- (1-Hydroxy-1-methylethyl) phenyl] prop-2-inyl} isoindole-1,3-dione

This compound was prepared in an analogous manner. to that of preparation 10 replacing the 2,2,2-trifluoro-1- (4-iodophenyl) ethanol  by 2- (3-iodophenyl) propan-2-ol (preparation 25). MS (m / e,%): 319 (M +, 90), 301 (80), 160 (100)

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Preparation 27

2- {3- [3- (1-Hydroxy-1-methylethyl) phenyl] propyl} isoindole-1,3-dione

This compound was prepared in an analogous manner. to that of preparation 11 replacing the 2- {3- [4- (2,2,2-Trifluoro-1-hydroxyethyl) phenyl] prop-2-inyl} isoindole-1,3-dione (preparation 10) by 2- {3- [3- (1-hydroxy-1-methylethyl) phenyl] prop-2-inyl} isoindole-1,3-dione  (preparation 26). MS (m / e,%): 305 (M + - H2O, 80), 145 (100).

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Preparation 28

2- [3- (3-Aminopropyl) phenyl] propan-2-ol

This compound was prepared in an analogous manner. to that of preparation 12 replacing the 2- {3- [4- (2,2,2-Trifluoro-1-hydroxyethyl) phenyl] propyl} isoindole-1,3-dione  by 2- {3- [3- (1-hydroxy-1-methylethyl) phenyl] propyl} isoindole-1,3-dione (preparation 27). MS (m / e,%): 193 (M +, 30), 162 (60), 145 (100)

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Preparation 29

4- [3- (1,3-Dioxo-1,3-dihydroisoindol-2-yl) prop-1-inyl] benzonitrile

This compound was prepared in an analogous manner. to that of preparation 10 using the starting materials appropriate. 1 H-NMR (CDCl 3) δ: 7.8 (m, 6H), 7.6 (d, 2H), 4.6 (s, 2H). MS (m / e,%): 286 (M +, 100).

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Preparation 30

4- [3 - [(1,3-Dioxo-1,3-dihydroisoindol-2-yl) propyl] benzonitrile

This compound was prepared in an analogous manner. to that of preparation 11 replacing the 2- {3- [4- (2,2,2-Trifluoro-1-hydroxyethyl) phenyl] prop-2-inyl} isoindole-1,3-dione  (preparation 10) by 4- [3- (1,3-Dioxo-1,3-dihydroisoindol-2-yl) prop-1-inyl] benzonitrile   (preparation 29). 1 H-NMR (CDCl 3) δ: 7.8 (dd, 2H), 7.7 (dd, 2H), 7.5 (d, 2H), 7.3 (d, 2H), 3.7 (t, 2H), 2.7 (t, 2H), 2.0 (m, 2H). MS (m / e,%): 290 (M +, 60), 161 (100).

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Preparation 31

4- (3-Aminopropyl) benzonitrile

This compound was prepared in an analogous manner. to that of preparation 12 replacing 2- {3- [4- (2,2,2-Trifluoro-1-hydroxyethyl) phenyl] propyl} isoindole-1,3-dione  by 4- [3 - [(1,3-dioxo-1,3-dihydroisoindol-2-yl) propyl] benzonitrile (preparation 30). 1 H-NMR (CDCl 3) δ:  7.5 (d, 2H), 7.3 (d, 2H), 2.7 (m, 4H), 1.8 (m, 2H). MS (m / e,%): 160 (M +, 20), 143 (100).

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Preparation 32

2-Chloro-3,4-dimethoxybenzaldehyde oxime

A mix of 2-Chloro-3,4-dimethoxybenzaldehyde (1.5 g, 7.5 mmol), hydroxylamine hydrochloride (650 mg, 9.4 mmol), and sodium acetate (1.5 g, 18.8 mmol) in 30 ml of MeOH and 15 ml of water was heated at 65 ° C for 18 hours. The mixture is diluted with water and extracted with EtOAc. The organic extract was washed successively with water and brine, dried over MgSO4, and concentrated to provide 1.7 g of a solid. 1 H-NMR (CDCl 3) δ: 8.5 (s, 1H), 7.6 (d, 1H), 6.8 (d, 1H), 3.9 (s, 3H), 3.8 (s, 3H). MS (m / e,%): 215 (M +, 40), 199 (100).

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Preparation 33

2-Chloro-3,4-dimethoxybenzylamine

To a stirred solution of the product of the Preparation 32 (1.7 g, 7.8 mmol) in 40 ml of THF at 0 ° C was added slowly drop by drop a 1 M solution of LAH in THF (17 ml, 17 mmoles). The mixture was allowed to slowly warm to RT after heated to reflux for 2 hours. The mixture was cooled to 0 ° C and The reaction was quenched with MeOH added slowly drop by drop. The The mixture was diluted with water and extracted with chloroform. Emulsion resulting was filtered through diatomaceous earth and separated The filtered phases. The organic extract was washed with water, dried (MgSO 4) and concentrated to provide 1.1 g of a oil. MS (m / e,%): 202 (M +, 100).

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Preparation 3. 4

3-Ethoxy-4-methoxybenzaldehyde oxime

This compound was prepared in an analogous manner. to that of preparation 32 using the starting materials appropriate. 1 H-NMR (CDCl 3) δ: 8.1 (s, 1H), 7.2 (d, 1H), 7.0 (m, 1H), 6.8 (d, 1H), 4.1 (c, 2H), 3.9 (s, 3H), 1.5 (t, 3H). MS (m / e,%): 195 (M +, 100).

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Preparation 35

3-Ethoxy-4-methoxybenzylamine

This compound was prepared in an analogous manner. to that of preparation 33 using the starting materials appropriate. 1 H-NMR (CDCl 3) δ: 6.9 (s, 1H), 6.8 (s, 2H), 4.1 (c, 2H), 3.8 (s, 2H), 1.5 (t, 3H). EM (m / e,%): 181 (M +, 100).

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Preparation 36

2- (3-Pyridin-4-ylpropyl) isoindole-1,3-dione

To a stirred solution of the product of 4-pyridimpropanol (2.0 g, 14.5 mmol), phthalimide (2.1 g, 14.5 mmol), and triphenylphosphine (4.9 g, 15.2 mmol) in 50 ml of THF at 0 ° C was added dropwise diethyl azodicarboxylate (2.5 ml, 16.0 mmol). The mixture was allowed to warm slowly until TA was then stirred overnight. The mixture was diluted with 0.1 N hydrochloric acid and washed with diethyl ether. The extract Aqueous was basified with 6 N sodium hydroxide and extracted with EtOAc. The organic extract was washed with 1 N sodium hydroxide and water, dried over MgSO4, and concentrated to provide 1.8 g of a solid. 1 H-NMR (CDCl 3) δ: 8.5 (s, 2H), 7.8 (m, 2H), 7.7 (m, 2H), 7.2 (m, 2H), 3.7 (t, 2H), 2.7 (t, 2H), 2.0 (m, 2H).

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Preparation 37

3-Pyridin-4-ylpropylamine

This compound was prepared in an analogous manner. to that of preparation 12 using the title compound of the Preparation 36. 1 H-NMR (CDCl 3) δ: 8.4 (m, 2H), 7.1 (m, 2H), 2.7 (t, 2H), 2.6 (t, 2H). MS (m / e,%): 136 (M +, 30), 119 (35), 107 (100).

The compounds of formula (I) were prepared as It is described in the following examples.

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Example one

N 4 - (3,5-Dimethoxybenzyl) -N 2 - (2-pyridin-2-yl-ethyl) pyrido [2,3-d] pyrimidin-2,4-diamine

A mixture of the title compound of the Preparation 1 (174 mg, 0.5 mmol), 2- (2-aminoethyl) -pyridine (386 mg, 3.2 mmol), and DIPEA (0.2 ml, 1.1 mmol) in 1 ml of DMSO was heated to 90 ° C for 18 hours. The mixture was poured into water and extracted with methylene chloride The organic extract was washed with brine, dried over MgSO4 and concentrated. The resulting residue is triturated with EtOAc to give 83 mg (40%) of the title compound, m.p. 157-8 ° C. 1 H-NMR (DMSO-d6): δ 8.6 (dd, 1H), 8.5 (d, 1H), 8.4 (dd, 1H), 7.7 (m, 1H), 7.2 (m, 1H), 7.0 (m, 2H), 6.5 (m, 2H), 6.3 (m, 1H), 4.6 (m, 2H), 3.7 (s, 6H), 3.6 (m, 2H), 3.0 (m, 2H). EM (m / z,%): 417 (100). Analysis calculated for C 23 H 24 N 6 O 2: C, 66.3; H, 5.8; N, 20.2. Found: C, 65.3; H, 6.3; N, 18.3. The compounds of the Examples 2 to 5 were prepared analogously to that described in the Example 1 using the appropriate starting materials.

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Example 2

N 4 - (3,5-Dimethoxybenzyl) -N 2 - (2-pyridin-3-yl-ethyl) pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 185-6 ° C. Calculated analysis for C 23 H 24 N 6 O 2: C, 66.3; H, 5.8; N, 20.2. Found: C, 64.5; H, 5.6; N, 19.6.

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Example 3

N 4 - (3,5-Dimethoxybenzyl) - N 2 - (2-pyridin-4-ylethyl) -pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 234-5 ° C. Calculated analysis for C 23 H 24 N 6 O 2: C, 66.3; H, 5.8; N, 20.2. Found: C, 66.0; H, 5.6; N, 20.0.

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Example 4

N 4 - (3,5-Dimethoxybenzyl) - N 2 -2-pyridin-3-ylmethylpyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 222-3 ° C. Calculated analysis for C 22 H 22 N 6 O 2: C, 65.7; H, 5.5; N, 20.9. Found: C, 65.5; H, 5.5; N, 20.8.

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Example 5

N 2 -N 4 -Bis- (3,5-dimethoxybenzyl) pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 159-160 ° C. Analysis calculated for C 25 H 27 N 5 O 4: C, 65.1; H, 5.9; N, 15.2. Found: C, 65.3; H, 5.9; N, 15.1.

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Example 6

N 4 - (3,5-Dimethoxybenzyl) - N 2 - [2- (4-methoxyphenyl) ethyl] pyrido [2,3-d] pyrimidin-2,4-diamine

A mixture of the title compound of preparation 1 (100 mg, 0.5 mmol), p -methoxyphenethylamine (89 µL, 0.61 mmol), and DIPEA (1.05 µL, 0.61 mmol) at 0 , 8 ml of DMSO was heated at 90 ° C for 18 hours. The compound was isolated and purified from the crude reaction mixture by direct injection in preparative reverse phase HPLC (Shimadzu Corp .; Kyoto, Japan) using a gradient in acetonitrile / water steps containing 0.1% ammonium hydroxide as eluent. Fractions containing the desired product were combined, concentrated and the residue recrystallized from IPA to give 80 mg (60%) of the title compound, mp 88-9 ° C. Analysis calculated for C 25 H 27 N 5 O 3: C, 67.4; H, 6.1; N, 15.7. Found: C, 67.0; H, 6.3; N, 15.2.

The compounds of examples 7 to 31 are prepared analogously to that described in example 6 using the appropriate starting materials. Exceptions are specified specific to the reaction and / or purification conditions employed

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Example 7

N 4 - (3,5-Dimethoxybenzyl) - N 2 - (3-phenylpropyl) pyrido [2,3-d] pyrimidin-2,4-diamine

1 H-NMR (DMSO-d6): 8.6 (dd, 1H), 8.4 (d, 1H), 7.2 (m, width, 3H), 7.1 (m, width, 2H), 7.0 (m, width, 1H), 6.5 (s, width, 2H), 6.3 (s, width, 1H), 4.6 (m, width, 2H), 3.7 (s, width, 6H), 2.6 (m, width, 2H), 1.8 (m, width, 2H), 1.7 (m, width, 2H). MS (m / z, %): 417 (100). MS (m / z,%): 430 (100).

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Example 8

N 2 - [2- (4-Chlorophenyl) ethyl] - N 4 - (3,5-dimethoxybenzyl) pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 85-90 ° C. Calculated analysis for C 24 H 24 N 5 O 2 Cl: C, 64.1; H, 5.4; N, 15.6. Found: C, 63.6; H, 5.7; N, 15.3.

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Example 9

N 2 -Benzyl- N 4 - (3,5-dimethoxybenzyl) pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 84-85 ° C. Calculated analysis for C 23 H 23 N 5 O 2: C, 68.8; H, 5.8; N, 17.4. Found: C, 68.5; H, 6.0; N, 17.4.

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Example 10

N 4 - (3,5-Dimethoxybenzyl) - N 2 - (2-thiophen-2-yl-ethyl) pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 120-1 ° C. Calculated analysis for C 22 H 23 N 5 O 2 S: C, 62.7; H, 5.5; N, 16.6. Found: C, 62.2; H, 6.0; N, 15.6.

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Example eleven

2- (4 - {[4- (3,5-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] methyl} -phenyl) propan-2-ol

m.p. 105-9 ° C. Calculated analysis for C 26 H 29 N 5 O 3: C, 68.0; H, 6.4; N, 15.2. Found: C, 67.7; H, 6.6; N, 14.4.

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Example 12

N 4 - (3,5-Dimethoxybenzyl) - N 2 -2-phenethylpyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 112-4 ° C. Calculated analysis for C 24 H 25 N 5 O 2: C, 69.4; H, 6.1; N, 16.9. Found: C, 68.5; H, 6.9; N, 15.3.

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Example 13

N 4 - (3,5-Dimethoxybenzyl) - N 2 - [2- (3,5-dimethoxyphenyl) ethyl] pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 95-100 ° C. Analysis calculated for C 26 H 29 N 5 O 4: C, 65.7; H, 6.2; N, 14.7 Found: C, 65.5; H, 6.6; N, 14.5.

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Example 14

N 4 - (3,4-Dimethoxybenzyl) - N 2 - [2- (3-fluorophenyl) ethyl] pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 227-8 ° C. Calculated analysis for C 24 H 24 N 5 O 2 F: C, 66.5; H, 5.6; N, 16.2. Found: C, 66.6; H, 5.6; N, 16.2.

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Example fifteen

N 4 - (3,4-Dimethoxybenzyl) - N 2 - [2- (2-fluorophenyl) ethyl] pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 220-2ºC. Calculated analysis for C 24 H 24 N 5 O 2 F: C, 66.5; H, 5.6; N, 16.2. Found: C, 66.4; H, 5.5; N, 16.1.

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Example 16

N 4 - (3,4-Dimethoxybenzyl) -N 2 - [2- (4-fluorophenyl) ethyl] pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 225-6 ° C. Calculated analysis for C 24 H 24 N 5 O 2 F: C, 66.5; H, 5.6; N, 16.2. Found: C, 66.4; H, 5.6; N, 16.2.

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Example 17

N 4 - (3,4-Dimethoxybenzyl) - N 2 -phenethylpyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 217-8 ° C. Calculated analysis for C 24 H 25 N 5 O 2: C, 69.4; H, 6.1; N, 16.9. Found: C, 69.4; H, 6.0; N, 16.8.

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Example 18

N 4 - (3,4-Dimethoxybenzyl) - N 2 - (4-phenylbutyl) pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 194-5 ° C. Calculated analysis for C 26 H 29 N 5 O 2: C, 70.4; H, 6.6; N, 15.8. Found: C, 70.5; H, 6.6; N, 15.9.

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Example 19

N 4 - (3,4-Dimethoxybenzyl) - N 2 - (2-phenoxyethyl) pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 227-8 ° C. Calculated analysis for C 24 H 25 N 5 O 3: C, 66.8; H, 5.8; N, 16.2. Found: C, 66.7; H, 5.7; N, 16.2.

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Example twenty

N 4 - (3,4-Dimethoxybenzyl) - N 2 - (2-trifluoromethylbenzyl) pyrido [2,3-d] pyrimidin-2,4-diamine

m.p. 227-8 ° C. Calculated analysis for C 24 H 22 N 5 O 2 F 3: C, 61.4; H, 4.7; N, 14.9. Found: C, 60.6; H, 4.9; N, 14.8.

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Example twenty-one

2- (4- {3- [4- (3,4-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] propyl} phenyl) propan-2-ol

The preparative HPLC fractions in phase inversely containing the desired crude product were concentrated and The residue was recrystallized from EtOAc. m.p. 198-9 ° C. Analysis calculated for C 28 H 33 N 5 O 3: C, 69.0; H 6.8; N, 14.4. Found: C, 68.7; H, 6.9; N, 14.3.

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Example 22

2- (4- {3- [4- (3,5-Dimethoxybenzylamino) pyrimidin-2-ylamino-2-ylamino] -propyl} phenyl) propan-2-ol

(amorphous solid) m.p. 70-5ºC. 1 H-NMR (DMSO-d 6): 8.6 (dd, 1H), 8.4 (d, 1H), 7.3 (m, width, 2H), 7.1 (m, width, 2H), 7.0 (m, width, 2H), 6.5 (s, width, 2H), 6.3 (s, width, 1H), 4.6 (m, width, 2H), 3.7 (s, width, 6H), 2.6 (m, width, 2H), 1.8 (m, width, 2H), 1.7 (m, width, 2H), 1.4 (s 6H). MS (m / z,%): 430 (100). MS (m / z,%): 488 (M + +1, 100), 470 (50). Analysis calculated for C 28 H 33 N 5 O 3: C, 69.0; H, 6.8; N, 14.4. Found: C, 67.1; H, 7.0; N, 12.1.

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Example 2. 3

1- (4 - {[4- (3,4-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] methyl} phenyl) -2,2,2-trifluoroethanol

The preparative HPLC fractions in phase inversely containing the desired crude product were concentrated and The residue was recrystallized from acetonitrile / water. m.p. 121-2 ° C. Analysis calculated for C 25 H 24 N 5 O 3 F 3: C, 60.1; H, 4.8; N, 14.0. Found: C, 58.0; H, 5.0; N, 13.4.

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Example 24

1- (4- {3- [4- (3,4-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] -propyl} phenyl) ethanone

The reaction mixture that contained the product Gross was poured into water and the resulting precipitate was removed by filtration. The residue was dissolved in 30 ml of MeOH and 10 was added ml of 1 N hydrochloric acid. The mixture was stirred at temperature ambient for four hours, concentrated, neutralized with a 5% sodium bicarbonate solution and extracted with dichloromethane The organic extracts were combined and washed with brine, dried over MgSO4 and concentrated. He crude product was purified by reverse phase HPLC as described in example 6. The fractions containing the desired product was concentrated and the residue recrystallized from acetonitrile / water to provide a solid, m.p. 184-5 ° C. Analysis calculated for C 27 H 29 N 5 O 3: C, 68.8; H, 6.2; N, 14.9. Found: C, 68.7; H, 6.0; N, 14.6.

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Example 25

1- (4- {3- [4- (3,4-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] propyl} -phenyl) -2,2,2-trifluoroethanol

m.p. 222-4 ° C. 1 H-NMR (DMSO-d 6): δ 8.6 (dd, 1H), 8.4 (dd, 1H), 7.4 (m, width, 2H), 7.3 (m, 1H), 7.0 (m, width, 2H), 6.8 (s, width, 2H), 6.7 (d, 1H), 5.0 (m, width, 1H), 4.6 (s, width, 2H), 3.7 (m, 6H), 3.3 (m, 2H), 2.6 (m, width, 2H), 1.8 (m, width, 2H). MS (m / z,%): 528 (M +, 100).

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Example 26

N 2 -3- (Benzo [1,2,5] oxadiazol-5-yl-propyl) - N 4 - (3,4-dimethoxy-benzyl) pyrido [2,3-d] pyrimidin -2,4-diamine

The product was isolated by gel chromatography of silica eluting with 2.5% MeOH in 2N ammonia / dichloromethane to give a gross solid. Crushing with EtOAc provided a solid, m.p. 191-3 ° C. 1 H-NMR (DMSO-d6): δ 8.6 (dd, 1H), 8.4 (dd, 1H), 7.9 (m, width, 1H), 7.7 (m, 1H), 7.5 (m, width, 1H), 7.0 (m, width, 2H), 6.8 (m, 2H), 4.6 (m, width, 2H), 3.7 (m, 6H), 3.4 (m, 2H), 2.7 (m, width, 2H), 1.9 (m, width, 2H). MS (m / z,%): 472 (M +, 100).

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Example 27

N 2 -3- (Benzothiazol-6-yl-propyl) - N 4 - (3,4-dimethoxybenzyl) pyrido [2,3-d] pyrimidin-2,4-diamine

The product was isolated by gel chromatography of silica eluting with 2.5% MeOH in 2 N ammonia / dichloromethane to give a solid, m.p. 191-3 ° C. MS (m / z,%): 487 (M +, 100). Analysis calculated for C 26 H 26 N 6 O 2 S: C, 64.2; H, 5.4; N, 17.3. Found: C, 64.5; H, 5.5; N, 16.8.

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Example 28

N 4 - (3,4-Dimethoxybenzyl) - N 2 - {3- [3- (2-methyl- [1,3] -dioxolan-2-yl) phenyl] propyl} pyrido [2 , 3-d] pyrimidin-2,4-diamine

1 H-NMR (DMSO-d6): δ 8.6 (d, 1H), 8.3 (d, 1H), 7.2 (m, width, 3H), 7.0 (m, width, 3H), 6.9 (m, width, 2H), 4.6 (m, width, 2H), 4.1 (m, 2H), 3.9 (m, 2H), 3.7 (m, width, 8H), 2.6 (m, width, 2H), 1.8 (m, width, 21-1) 1.5 (s, 3H). EM (m / z,%): 516 (M + +1, 100).

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Example 29

2- (3- {3- [4- (3,4-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] propyl} phenyl) propan-2-ol

m.p. 204-5 ° C. 1 H-NMR (DMSO-d 6): δ 8.6 (d, 1H), 8.3 (d, 1H), 7.3 (m, width, 1H), 7.2 (m, width, 1H), 7.1 (m, width, 1H), 7.0 (m, width, 3H), 6.8 (s, width, 2H), 4.6 (s, width, 2H), 3.7 (m, width, 6H), 3.3 (m, width, 2H), 2.6 (m, width, 2H), 1.8 (m, width, 2H), 1.4 (s, 6H). MS (m / z,%): 488 (M + + 1, 100).

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Example 30

4- {3- [4- (3,4-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] propyl} benzonitrile

m.p. 220-2ºC. 1 H-NMR (DMSO-d 6): δ 8.6 (d, 1H), 8.3 (d, 1H), 7.7 (m, width, 2H), 7.4 (m, width, 2H), 7.0 (m, width, 2H), 6.8 (s, width, 2H), 4.6 (s, width, 2H), 3.7 (m, width, 6H), 3.3 (m, width, 2H), 2.7 (m, width, 2H), 1.8 (m, width, 2H). MS (m / z,%): 455 (M + + 1,100).

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Example 31

N 4 - (3,5-Dimethoxybenzyl) - N 2 - (3-pyridin-4-yl-propyl) pyrido [2,3-d] pyrimidin-2,4-diamine

1 H-NMR (CD 2 Cl 2): δ 9.1 (d, 1H), 8.9 (m, width, 1H), 8.4 (m, width, 3H), 7.2 (m, width, 2H), 7.1 (m, width, 1H), 7.0 (s, width, 1H), 6.9 (d, 1H), 6.6 (d, 1H), 4.8 (d, 2H), 3.7 (s, 3H), 3.7 (s, 3H), 3.6 (m, 2H), 2.7 (m, 2H), 2.0 (m, 2H). MS (m / z,%): 432 (M + + 1, 20), 256 (15), 237 (30), 216 (100).

The compounds of the examples 32-35 were prepared according to the following generalized procedure The exceptions to the conditions used for reaction and / or purification.

To a stirred solution of 2,4-dichloro-pyrido [2,3-d] pyrimidine (0.75 mmol) and DIPEA (1.5 mmol) in 3 ml DMSO 0.75 mmol were added of an amine corresponding to formula (V), scheme 1, STEP 1, to room temperature. The mixture was stirred at room temperature. for one hour, and then 2.25 mmol of an amine was added corresponding to formula (VII), Scheme 1, STAGE 2, and DIPEA additional (2.25 mmol), and the mixture was heated at 90 ° C for two hours.

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Example 32

N 4 - (3,4-Dimethoxybenzyl) - N 2 - (3-phenylpropyl) pyrido [2,3-d] pyrimidin-2,4-diamine

The compound was isolated and purified from the crude reaction mixture by direct injection into an HPLC preparatory in reverse phase using a gradient in stages of acetonitrile / water containing 0.1% ammonium hydroxide as eluent Fractions containing the compound were combined and concentrated to give a solid, m.p. 190-2 ° C. 1 H-NMR (d 6 -DMSO): δ 8.6 (dd, 1H), 8.4 (dd, 1H), 7.2 (m, width, 3H), 7.1 (m, width, 2H), 7.0 (m, width, 2H), 6.8 (m, width, 2H), 4.6 (m, width, 2H), 3.7 (d, width, 6H), 3.3 (m, width, 2H), 2.6 (m, width, 2H), 1.8 (m, 2H). EM (m / e,%): 431 (M + + 1, 50), 430 (M +, 100).

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Example 33

N 4 - (3,5-Dimethoxybenzyl) - N 2 - (3-phenoxyethyl) pyrido [2,3-d] pyrimidin-2,4-diamine

The compound was isolated and purified from the crude reaction mixture by direct injection into an HPLC preparatory in reverse phase using a gradient in stages of acetonitrile / water containing 0.1% ammonium hydroxide as eluent Fractions containing the compound were combined, they were concentrated, and the residue recrystallized from IPA / water. m.p. 165-7 ° C. 1 H-NMR (d6 -DMSO): 8.6 (dd, 1H), 8.4 (d, 1H), 7.2 (m, width, 3H), 7.1 (m, width, 2H), 7.0 (m, 1H), 6.9 (m, width, 2H), 6.8 (m, width, 1H), 6.5 (d, 2H), 6.4 (s, 1H), 4.6 (m, width, 2H), 4.0 (m, width, 2H), 3.7 (m, width, 8H). MS (m / e,%): 433 (M + + 1, 50), 432 (M +, 100).

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Example 3. 4

N 4 - (3-Ethoxy-4-methoxybenzyl) - N 2 - (3-phenylpropyl) pyrido [2,3-d] pyrimidin-2,4-diamine

The compound was isolated and purified from the crude reaction mixture by direct injection into an HPLC preparatory in reverse phase using a gradient in stages of acetonitrile / water containing 0.1% ammonium hydroxide as eluent Fractions containing the compound were combined, they were concentrated, and recrystallized from acetonitrile / water. m.p. 181-2 ° C. Analysis calculated for C 26 H 29 N 5 O 2: C, 70.4; H, 6.6; N, 15.8. Found: C, 70.7; H, 6.9; N, 15.9.

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Example 35

2- (4- {3- [4- (3-Ethoxy-4-methoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] -propyl} phenyl) propan-2-ol

The compound was isolated and purified from the crude reaction mixture by direct injection into an HPLC preparatory in reverse phase using a gradient in stages of acetonitrile / water containing 0.1% ammonium hydroxide as eluent Fractions containing the desired compound were combined, concentrated, and recrystallized from acetonitrile / water m.p. 150-2ºC. Calculated analysis for C 29 H 35 N 5 O 3: C, 69.4; H, 7.0; N, 13.7. Found: C, 69.1; H, 6.9; N, 13.7.

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Example 36

1- (4- {3- [4- (3,4-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] propyl} -phenyl) -2,2,2-trifluoroethanone

Was added 1,1,1-triacetoxy-1,1-dihydro-1,2-benciodoxol-3 (1H) -one (180 mg, 0.43 mmol) to a stirred solution of the compound of title of example 25 (150 mg, 0.28 mmol) in 15 ml dichloromethane at room temperature. The mixture was stirred at room temperature. for six hours, then an additional portion of 1,1,1-triacetoxy-1,1-dihydro-1,2-benciodoxol-3 (1H) -one  (180 mg, 0.43 mmol) and the mixture was stirred at room temperature overnight. The mixture was diluted with chloroform, washed with water, dried over MgSO4 and concentrated to give an oil. The silica gel chromatography eluting with 5% 2N MeOH in ammonia / dichloromethane provided an oil. Crushing with EtOAc provided 45 mg of a solid, m.p. 198-200 ° C. 1 H-NMR (d_ {6} -DMSO): δ 8.6 (dd, 1H), 8.4 (dd, 1H),  7.9 (m, width, 2H), 7.5 (m, width, 2H), 7.0 (m, width, 2H), 6.8 (m, width, 2H), 4.6 (m, 2H), 3.7 (m, 6H), 3.3 (m, 2H), 2.7 (m, width, 2H), 1.9 (m, width, 2H). MS (m / e,%): 526 (M +, 100).

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Example 37

1- (3- {3- [4- (3,4-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] propyl} phenyl) ethanone

A mixture of the title compound of the Preparation 27 (419 mg, 0.81 mmol) in 8 ml of THF containing 10 ml of 6 N hydrochloric acid was stirred at room temperature for six hours. The reaction mixture was extracted with EtOAc, the extract Organic was washed with 5% sodium bicarbonate solution, and then concentrated to give a solid, m.p. 152-5 ° C. 1 H-NMR (d 6 -DMSO): 8.8 (d, 1H), 8.7 (d, 1H), 7.7 (m, 2H), 7.4 (m, 3H), 7.0 (s, 1H), 6.8 (m, 2H), 4.6 (d, 2H), 3.7 (s, 3H), 3.6 (s, 3H), 3.5 (m, 2H), 2.7 (m, 2H), 2.5 (s, 3H), 1.9 (m, 2H). MS (m / e,%): 471 (M +, 30), 470 (100)

Biological methodologies PDE 2 enzyme isolation

PDE 2 enzyme was isolated from human thrombocytes with approximately 1.4 l of blood from multiple donors employed to prepare the sediments of the thrombocytes. The thrombocytes are resuspended in approximately 75 ml of lysis buffer [Tris 20 mM pH 7.2, 5 mM MgCl 2, 250 mM sucrose, 1 mM DTT, 1 µl / 2 ml of Sigma protease inhibitor No. 8340; Sigma-Aldrich; St. Louis, MO] and were lysed by ultrasound at 4 ° C, with 3 cycles of 1 minute of impulses and then centrifuged at 4 ° C overnight at 100,000 x g. Lysates rinses were loaded into an AKLC Explorer FPLC (Amersham Biosciences; Piscataway, NJ) in a series of three separations chromatographic, with an average loading volume of 25 ml. One was used 5 ml HiTrap Q anion exchange column (Amersham Biosciences). A buffer [20 mM Tris pH 7.2, MgCl 2 5 was mixed mM, 1 µl / 2 ml of Sigma protease inhibitor No. 8340] with a buffer gradient B [20 mM Tris pH 7.2, 500 mM NaCl, MgCl 2 5 mM, 1 µl / 2 ml of Sigma protease inhibitor No. 8340] in 20 column volumes from an initial concentration of buffer B of 0% to a final concentration of 100%. The maximum values that hydrolyse cGMP were scored with an average resolution at low salt concentration-125 mM NaCI (PDE 5) and High salt concentration-325 mM NaCI (PDE 2). The two main fractions of cGMP activity (PDE 5 and PDE 2) are isolated and combined separately. The total fraction combined PDE 2 was approximately 40 ml, which was dispensed in cryovials  of 200 µl / vial and deposited in storage at -80 ° C.

PDE 2 Enzyme Binding Assay

The inhibitory activity of the compounds of formula (I) on recombinant or isolated PDE 2, and other PDE, is determined using the scintillation proximity test kits [^ 3 H] cAMP (SPA) of Amersham International (Little Chalfont, England). SPA trials were performed using 96 well plates. Itium SPA silicate pearls with PDE (Amersham Biosciences) preferentially bound the nucleotide linear, GMP, compared to the cyclic nucleotide, cGMP. Was added 3 H-cGMP to the reaction and when the product, 3 H-GMP, was in close proximity to the pearl, the twinkling inside the pearl was excited, which detected using a packard scintillation counter (Perkin-Elmer Life Sciences; Boston, MA). The enzyme concentration used was in the linear range and was determined the K m of the enzyme (15 µM). Concentration substrate end was <1/3 of the K_ {m} (1 µM) so that the IC_ {50} values would approximate the K_ {i} values. The assay was validated using the compounds of the literature as controls before testing the compounds. Then they were compared the measurements of the catalytic activity of PDE obtained in the presence of the test compound, and those obtained in the absence of test compound, and the IC50 value was determined.

The radioactive substrates and the products of the PDE reaction was determined quantitatively using a liquid scintillation counter RACK-BETA 1219 (LKB Wallac; Freiburg, FRG). IC 50 values (concentrations with 50% inhibition) were determined with cAMP or 1 µM cGMP using the maximum value fractions. The data was adjusted with four parameters with the help of a logistics function sigmoidal

Using the PDE 2 enzyme described previously isolated from human thrombocytes and the procedure to test the Test compounds for enzyme inhibition, it was determined an IC 50 of 1.7 µM for EHNA. In addition, it was also determined an IC 50 of 3 nM for the 9- (1-acetyl-4-phenyl-butyl) -2- (3,4-dimethoxy-benzyl) -1,9-dihydropurin-6-one  using the procedure mentioned above.

The compounds of formula (I) have generally an inhibitory activity, expressed as IC50 compared to PDE 2, which is <1,000 nM. The intervals of the PDE 2 inhibitory activity for compounds of formula (I) in examples 1-37 are set out in the table one.

TABLE 1

7

The capacity of PDE 2 inhibitors, including compounds of formula (I), to treat the fracture and / or bone defect, or promote bone growth, can be Demonstrate according to the following protocols.

Model of femoral transverse fracture in rats

Male Sprague-Dawley rats 3 to 4 months old were used. The animals were anesthetized with ketamine and xylazine at doses of 100 and 10 mg / kg, respectively. The right leg of each rat was shaved and cleaned. A 1 cm incision was made just lateral to the knee and the femoral condyle was exposed. A Kirschner cable (1.14 mm in diameter) was introduced into the intramedullary canal through the intercondylar portion, to serve with internal stabilization. The muscle incision was closed with vicrile and the skin incision was closed with stainless steel wound clips. The mean diaphysis of the fixed femur was fractured by a three-point flexure device directed by a hanging weight. The rats were allowed to carry all the weight and unrestricted activity after waking from anesthesia. Test compounds were administered on several days after surgery by percutaneous injection over the fracture site. Animals were sacrificed after treatment and femurs were removed for analysis. Fracture healing was evaluated using radiographic, histological and biomechanical tests (F. Bonnarens, et al.), Journal of Orthopedic Research, 2 , 97-101 (1984).

Study protocol and results in the fracture model transverse femoral in rats

Three month old male rats underwent transverse fracture of the right femur with general anesthesia. A single dose (5 mg) of N 4 - (3,4-dimethoxy-benzyl) - N 2 - (3-phenyl-propyl) -pyrido [2,3-d] was injected percutaneously ] pyrimidin-2,4-diamine at fracture sites at the end of the fracture generation. Three weeks after the injection, the rats were sacrificed and the right femurs were extracted and analyzed. Fracture strength and stiffness (bone strength indices) increased by 19% and 62%, respectively, in femurs treated with N 4 - (3,4-dimethoxy-benzyl) - N 2 - (3-Phenyl-propyl) -pyrido [2,3-d] pyrimidin-2,4-diamine, compared with those of the femurs treated with placebo.

Periodic injection model in rats

Rats were anesthetized with isoflurane (2-3 minutes) in a localized driving chamber in an extraction hood. The right leg of each rat is He shaved and cleaned himself. A 0.5 mm diameter needle was preloaded coupled to a syringe with a formulation of the title compound for local injection. The formulation was injected into the subperiosteum of the femur in a volume of 5-15 µl for 14 days. The rats were sacrificed after dosing, collected the femurs, and then analyzed by radiography and dual X-ray densitometry (DEXA).

Study protocol and results in the injection model periodic in rats

The three-week-old male Sprague-Dawley male right femur received a daily injection of vehicle or test compound five times a week for two weeks. On day 15, all rats were sacrificed and the right femurs were removed for analysis. Periodic bone induction was evaluated using radiography and DEXA. Radiographs showed a new bone formation located on the injection site of all femurs treated with the test compound. The bone mineral content (BMC) of the injected region of the femur (area between the lesser trochanter and the center of the femur axis) was analyzed by DEXA comparing the rats treated with the test compound and those treated only with vehicle. In this model, N 4 - (3,5-dimethoxybenzyl) - N 2 - (2-pyridin-4-yl-ethyl) pyrido [2,3-d] pyrimidin-2,4-diamine , 2- (3- {3- [4- (3,4-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] propyl} phenyl) propan-2-ol and N 4 - ( 3,4-dimethoxybenzyl) - N 2 - (3-phenylpropyl) pyrido [2,3-d] pyrimidin-2,4-diamine increased the BMC by 34%, 49% and 33% respectively. In addition, 9- (1-acetyl-4-phenylbutyl) -2- (3,4-dimethoxybenzyl) -1,9-dihydropurin-6-one increased the BMC by 20% in the model mentioned above.

Claims (17)

1. A compound of formula (I) 8 and the salts of said compounds pharmaceutically acceptable, in the that: R 1 and R 2 are hydrogen or methoxy, with the condition that R1 and R2 are not both hydrogen or both methoxy; n is 1, 2, 3, or 4; X is a link; OR; S; C = O; -N (R) -; where R is hydrogen or (C 1 -C 3) alkyl; -C (OH) -; or -SO2; and Y is benzoxazolyl; benzothiazolyl; benzofurazanyl; benzofuranyl; benzothiadiazolyl; bencisoxazolyl; bencisothiazolyl; benzimidazolyl; pyridyl; isatinyl; oxindolyl; indazolyl; indolyl; phenyl; thienyl; or furanyl; where and is optionally independently substituted with between one and three halogen; trifluoromethyl; methoxy; -C (= O) CH 3; cyano; -C (CH 3) 2 OH; -CH (CH 3) OH; -CH (CF 3) OH; -C (C = O) CF 3; -SO2 NH2; -C (= O) OCH 3; -CH 2 COOH; 9 thiazolyl; or oxadiazolyl. 2. A compound of claim 1, in the that X is a bond, and Y is benzofurazanyl; thienyl; pyridyl; or phenyl, in which phenyl is optionally substituted independently with one or two halogen; trifluoromethyl; methoxy; -C (= O) CH 3; cyano; -C (CH 3) 2 OH; -CH (CH 3) OH; -CH (CF 3) OH; -C (C = O) CF 3; -SO2 NH2; -C (= O) OCH 3; -CH 2 COOH; thiazolyl; or oxadiazolyl. 3. A compound of claim 1 or 2, in that X is a bond, n is 2 or 3, and Y is thienyl; pyridyl; or phenyl, where phenyl is independently substituted with one or two methoxy; halogen; -C (CH 3) 2 OH; -CH (CF 3) OH; or -C (C = O) CF 3. 4. N 2, N 4 -bis- (3,5-Dimethoxybenzyl) pyrido [2,3-d] pyrimidin-2,4-diamine; N 4 - (3,5-Dimethoxybenzyl) - N 2 - (2-pyridin-4-ylethyl) pyrido [2,3-d] pyrimidin-2,4-diamine; N 4 - (3,5-Dimethoxybenzyl) - N 2 - (2-thiophene-2-ylethyl) pyrido [2,3-d] pyrimidin-2,4-diamine; N 4 - (3,5-Dimethoxybenzyl) - N 2 -2-phenethylpyrido [2,3-d] pyrimidin-2,4-diamine; N 4 - (3,5-Dimethoxybenzyl) - N 2 - [2- (3,5-dimethoxy-phenyl) ethyl] pyrido [2,3-d] pyrimidin-2,4-diamine; 2- (3- {3- [4- (3,4-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] -propyl} phenyl) propan-2-ol; N 4 - (3,4-Dimethoxybenzyl) - N 2 - [2- (4-fluorophenyl) ethyl] pyrido [2,3-d] pyrimidin-2,4-diamine; N 4 - (3,4-Dimethoxybenzyl) - N 2 -phenethylpyrido [2,3-d] pyrimidin-2,4-diamine; or N 4 - (3,4-Dimethoxybenzyl) - N 2 - (3-phenylpropyl) pyrido [2,3-d] pyrimidin-2,4-diamine; or a pharmaceutically acceptable salt of said compound. 5. A pharmaceutical composition comprising a compound of formula (I) of any of the claims 1-4, or a salt of said compound pharmaceutically acceptable, and a carrier, vehicle, or pharmaceutically diluent acceptable. 6. The use of a compound or salt according to any one of claims 1 to 4 in the manufacture of a medicine to treat a condition, disease or symptom mediated by PDE 2 in a mammal. 7. The use of claim 6, wherein said condition, disease, or symptoms is osteoporosis, hypertension  pulmonary, female sexual arousal disorder, memory or decreased cognition, platelet aggregation, angiogenesis vascular, dementia, cancer, arrhythmia, thrombosis, fracture and / or bone defect, delayed fracture or pseudoarthrosis, fusion of the spine, deep bone regrowth, reconstruction cranial facial, or hypoxia. 8. A method of claim 6, in which said condition is fracture and / or bone defect. 9. A pharmaceutical composition comprising a PDE 2 inhibitor according to formula (I) of the claim 1, a selective agonist of the EP2 receptor, and a pharmaceutically acceptable carrier, vehicle, or diluent. 10. A composition of claim 9, wherein said PDE 2 inhibitor is N 4 - (3,5-dimethoxybenzyl) - N 2 - (2-pyridin-4-ylethyl) pyrido [ 2,3-d] pyrimidin-2,4-diamine; 2- (3- {3- [4- (3,4-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] -propyl} -phenyl) -propan-2-ol; N 4 - (3,4-dimethoxybenzyl) - N 2 - (3-phenylpropyl) pyrido [2,3-d] pyrimidin-2,4-diamine; or a pharmaceutically acceptable salt of said compound. 11. A composition of claim 9 or 10, wherein said selective EP 2 receptor agonist is (3 - (((4- tert -butylbenzyl) - (pyridin-3-sulfonyl) amino) methyl) phenoxy ) acetic, or a pharmaceutically acceptable salt of said compound. 12. The use of any of the claims 6-8, which further includes administration to said mammal of a therapeutically effective amount of an agonist selective of the EP2 receptor; or a pharmaceutical composition that comprises a combination of said compound of formula (I) of the claim 1 and said selective receptor agonist EP2. 13. The use of claim 12, wherein said PDE 2 inhibitor is N 4 - (3,5-dimethoxybenzyl) - N 2 - (2-pyridin-4-ylethyl) pyrido [ 2,3-d] pyrimidin-2,4-diamine; 2- (3- {3- [4- (3,4-Dimethoxybenzylamino) pyrido [2,3-d] pyrimidin-2-ylamino] propyl} phenyl) propan-2-ol; N 4 - (3,4-dimethoxybenzyl) - N 2 - (3-phenylpropyl) pyrido [2,3-d] pyrimidin-2,4-diamine; or a pharmaceutically acceptable salt of said compound. 14. The use of claim 12 or 13, wherein said selective EP2 receptor agonist is (3 - (((4- tert -butylbenzyl) - (pyridin-3-sulfonyl) amino) methyl) phenoxy ) acetic, or a pharmaceutically acceptable salt of said compound. 15. The use of a PDE 2 inhibitor according with the formula (I) of claim 1 or a salt pharmaceutically acceptable of said inhibitor, in the manufacture of a medication for the treatment of a fracture and / or defect Bone in a mammal. 16. A compound or salt according to the claim 1 for use as a medicine. 17. A compound or salt according to the claim 1 for use as a medicament in the treatment of a  condition, disease or symptom mediated by PDE 2 in a mammal.